Inhibitors of glycogen synthase kinase 3

ABSTRACT

New pyrimidine or pyridine based compounds, compositions and methods of inhibiting the activity of glycogen synthase kinase (GSK3) in vitro and of treatment of GSK3 mediated disorders in vivo are provided. The methods, compounds and compositions of the invention may be employed alone, or in combination with other pharmacologically active agents in the treatment of disorders mediated by GSK3 activity, such as diabetes, Alzheimer&#39;s disease and other neurodegenerative disorders, obesity, atherosclerotic cardiovascular disease, essential hypertension, polycystic ovary syndrome, syndrome X, ischemia, traumatic brain injury, bipolar disorder, immunodeficiency or cancer.

CROSS-REFERENCES TO RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional Application No.60/230,480, filed Sep. 6, 2000, and is a continuation-in-part of U.S.application Ser. No. 09/336,038, filed Jun. 18, 1999 (now U.S. Pat. No.6,417,185), which claims the benefit of U.S. Provisional Application No.60/089,978, filed Jun. 19, 1998.

FIELD OF THE INVENTION

This invention relates to new pyrimidine and pyridine derivatives thatinhibit the activity of glycogen synthase kinase 3 (GSK3) and topharmaceutical compositions containing the compounds and to the use ofthe compounds and compositions, alone or in combination with otherpharmaceutically active agents. The compounds and compositions providedby the present invention have utility in the treatment of disordersmediated by GSK3 activity, such as diabetes, Alzheimer's disease andother neurodegenerative disorders, obesity, atheroscleroticcardiovascular disease, essential hypertension, polycystic ovarysyndrome, syndrome X, ischemia, especially cerebral ischemia, traumaticbrain injury, bipolar disorder, immunodeficiency and cancer.

BACKGROUND OF THE INVENTION

Glycogen synthase kinase 3 (GSK3) is a serine/threonine kinase for whichtwo isoforms, α and β, have been identified. Woodgett, Trends Biochem.Sci., 16:177-81 (1991). Both GSK3 isoforms are constitutively active inresting cells. GSK3 was originally identified as a kinase that inhibitsglycogen synthase by direct phosphorylation. Upon insulin activation,GSK3 is inactivated, thereby allowing the activation of glycogensynthase and possibly other insulin-dependent events, such glucosetransport. Subsequently, it has been shown that GSK3 activity is alsoinactivated by other growth factors that, like insulin, signal throughreceptor tyrosine kinases (RTKs). Examples of such signaling moleculesinclude IGF-1 and EGF. Saito et al., Biochem. J., 303:27-31 (1994);Welsh et al., Biochem. J. 294:625-29 (1993); and Cross et al., Biochem.J., 303:21-26 (1994).

Agents that inhibit GSK3 activity are useful in the treatment ofdisorders that are mediated by GSK3 activity. In addition, inhibition ofGSK3 mimics the activation of growth factor signaling pathways andconsequently GSK3 inhibitors are useful in the treatment of diseases inwhich such pathways are insufficiently active. Examples of diseases thatcan be treated with GSK3 inhibitors are described below.

Diabetes.

Diabetes mellitus is a serious metabolic disease that is defined by thepresence of chronically elevated levels of blood glucose(hyperglycemia). This state of hyperglycemia is the result of a relativeor absolute lack of activity of the peptide hormone, insulin. Insulin isproduced and secreted by the β cells of the pancreas. Insulin isreported to promote glucose utilization, protein synthesis, and theformation and storage of carbohydrate energy as glycogen. Glucose isstored in the body as glycogen, a form of polymerized glucose, which maybe converted back into glucose to meet metabolism requirements. Undernormal conditions, insulin is secreted at both a basal rate and atenhanced rates following glucose stimulation, all to maintain metabolichomeostasis by the conversion of glucose into glycogen.

The term diabetes mellitus encompasses several different hyperglycemicstates. These states include Type 1 (insulin-dependent diabetes mellitusor IDDM) and Type 2 (non-insulin dependent diabetes mellitus or NIDDM)diabetes. The hyperglycemia present in individuals with Type 1 diabetesis associated with deficient, reduced, or nonexistent levels of insulinthat are insufficient to maintain blood glucose levels within thephysiological range. Conventionally, Type 1 diabetes is treated byadministration of replacement doses of insulin, generally by a parentalroute. Since GSK3 inhibition stimulates insulin-dependent processes, itis consequently useful in the treatment of type 1 diabetes.

Type 2 diabetes is an increasingly prevalent disease of aging. It isinitially characterized by decreased sensitivity to insulin and acompensatory elevation in circulating insulin concentrations, the latterof which is required to maintain normal blood glucose levels. Increasedinsulin levels are caused by increased secretion from the pancreaticbeta cells, and the resulting hyperinsulinemia is associated withcardiovascular complications of diabetes. As insulin resistance worsens,the demand on the pancreatic beta cells steadily increases until thepancreas can no longer provide adequate levels of insulin, resulting inelevated levels of glucose in the blood. Ultimately, overt hyperglycemiaand hyperlipidemia occur, leading to the devastating long-termcomplications associated with diabetes, including cardiovasculardisease, renal failure and blindness. The exact mechanism(s) causingtype 2 diabetes are unknown, but result in impaired glucose transportinto skeletal muscle and increased hepatic glucose production, inaddition to inadequate insulin response. Dietary modifications are oftenineffective, therefore the majority of patients ultimately requirepharmaceutical intervention in an effort to prevent and/or slow theprogression of the complications of the disease. Many patients can betreated with one or more of the many oral anti-diabetic agentsavailable, including sulfonylureas, to increase insulin secretion.Examples of sulfonylurea drugs include metformin for suppression ofhepatic glucose production, and troglitazone, an insulin-sensitizingmedication. Despite the utility of these agents, 30-40% of diabetics arenot adequately controlled using these medications and requiresubcutaneous insulin injections. Additionally, each of these therapieshas associated side effects. For example, sulfonylureas can causehypoglycemia and troglitazone can cause severe hepatoxicity. Presently,there is a need for new and improved drugs for the treatment ofprediabetic and diabetic patients.

As described above, GSK3 inhibition stimulates insulin-dependentprocesses and is consequently useful in the treatment of type 2diabetes. Recent data obtained using lithium salts provides evidence forthis notion. The lithium ion has recently been reported to inhibit GSK3activity. Klein et al., PNAS 93:8455-9 (1996). Since 1924, lithium hasbeen reported to have antidiabetic effects including the ability toreduce plasma glucose levels, increase glycogen uptake, potentiateinsulin, up-regulate glucose synthase activity and to stimulate glycogensynthesis in skin, muscle and fat cells. However, lithium has not beenwidely accepted for use in the inhibition of GSK3 activity, possiblybecause of its documented effects on molecular targets other than GSK3.The purine analog 5-iodotubercidin, also a GSK3 inhibitor, likewisestimulates glycogen synthesis and antagonizes inactivation of glycogensynthase by glucagon and vasopressin in rat liver cells. Fluckiger-Isleret al., Biochem J 292:85-91 (1993); and Massillon et al., Biochem J299:123-8 (1994). However, this compound has also been shown to inhibitother serine/threonine and tyrosine kinases. Massillon et al., Biochem J299:123-8 (1994).

One of the main goals in the management of patients with diabetesmellitus is to achieve blood glucose levels as close to normal aspossible. In general, obtaining normal postprandial blood glucose levelsis more difficult than normalizing fasting hyperglycemia. In addition,some epidemiological studies suggest that postprandial hyperglycemia(PPHG) or hyperinsulinemia are independent risk factors for thedevelopment of macrovascular complications of diabetes mellitus.Recently, several drugs with differing pharmacodynamic profiles havebeen developed which target PPHG. These include insulin lispro, amylinanalogues, alpha-glucosidase inhibitors and meglitinide analogues.Insulin lispro has a more rapid onset of action and shorter duration ofefficacy compared with regular human insulin. In clinical trials, theuse of insulin lispro has been associated with improved control of PPHGand a reduced incidence of hypoglycemic episodes. Repaglinide, ameglitinide analogue, is a short-acting insulinotropic agent which, whengiven before meals, stimulates endogenous insulin secretions and lowerspostprandial hyperglycaemic excursions. Both insulin lispro andrepaglinide are associated with postprandial hyperinsulinaemia. Incontrast, amylin analogues reduce PPHG by slowing gastric emptying anddelivery of nutrients to the absorbing surface of the gut.Alpha-glucosidase inhibitors such as acarbose, miglitol and voglibosealso reduce PPHG primarily by interfering with thecarbohydrate-digesting enzymes and delaying glucose absorption. Yamasakiet al., Tohoku J Exp Med 1997 November; 183(3):173-83. The GSKinhibitors of the present invention are also useful, alone or incombination with the agents set forth above, in the treatment ofpostprandial hyperglycemia as well as in the treatment of fastinghyperglycemia.

Alzheimer's Disease.

GSK3 is also involved in biological pathways relating to Alzheimer'sdisease (AD). The characteristic pathological features of AD areextracellular plaques of an abnormally processed form of the amyloidprecursor protein (APP), so called β-amyloid peptide (β-AP) and thedevelopment of intracellular neurofibrillary tangles containing pairedhelical filaments (PHF) that consist largely of hyperphosphorylated tauprotein. GSK3 is one of a number of kinases that have been found tophosphorylate tau protein in vitro on the abnormal sites characteristicof PHF tau, and is the only kinase also demonstrated to do this inliving cells and in animals. Lovestone et al., Current Biology 4:1077-86(1994); and Brownlees et al., Neuroreport 8: 3251-3255 (1997).Furthermore, the GSK3 kinase inhibitor, LiCl, blocks tauhyperphosphorylation in cells. Stambolic et al., Current Biology6:1664-8 (1996). Thus GSK3 activity may contribute to the generation ofneurofibrillary tangles and consequently to disease progression.Recently it has been shown that GSK3β associates with another keyprotein in AD pathogenesis, presenillin 1 (PS1). Takashima et., PNAS95:9637-9641 (1998). Mutations in the PS1 gene lead to increasedproduction of β-AP, but the authors also demonstrate that the mutant PS1proteins bind more tightly to GSK3β and potentiate the phosphorylationof tau, which is bound to the same region of PS1.

Interestingly it has also been shown that another GSK3 substrate,β-catenin, binds to PS1. Zhong et al., Nature 395:698-702 (1998).Cytosolic β-catenin is targeted for degradation upon phosphorylation byGSK3 and reduced β-catenin activity is associated with increasedsensitivity of neuronal cells to β-AP induced neuronal apoptosis.Consequently, increased association of GSK3β with mutant PS1 may accountfor the reduced levels of β-catenin that have been observed in thebrains of PS1-mutant AD patients and to the disease related increase inneuronal cell-death. Consistent with these observations, it has beenshown that injection of GSK3 antisense but not sense, blocks thepathological effects of β-AP on neurons in vitro, resulting in a 24 hrdelay in the onset of cell death. Takashima et al., PNAS 90:7789-93.(1993). In these latter studies, the effects on cell-death are preceded(within 3-6 hours of β-AP administration) by a doubling of intracellularGSK3 activity, suggesting that genetic mechanisms may increase GSK3activity. Further evidence for a role for GSK3 in AD is provided by theobservation that the protein expression level (but, in this case, notspecific activity) of GSK3 is increased by 50% in postsynaptosomalsupernatants of AD vs. normal brain tissue. Pei et al., J NeuropatholExp 56:70-78 (1997). Thus, it is believed that specific inhibitors ofGSK3 will act to slow the progression of Alzheimer's Disease.

Other CNS Disorders

In addition to the effects of lithium described above, there is a longhistory of the use of lithium to treat bipolar disorder (manicdepressive syndrome). This clinical response to lithium may reflect aninvolvement of GSK3 activity in the etiology of bipolar disorder, inwhich case GSK3 inhibitors could be relevant to that indication. Insupport of this notion it was recently shown that valproate, anotherdrug commonly used in the treatment of bipolar disorder, is also a GSK3inhibitor. Chen et al., J. Neurochemistry 72:1327-1330 (1999). Onemechanism by which lithium and other GSK3 inhibitors may act to treatbipolar disorder is to increase the survival of neurons subjected toaberrantly high levels of excitation induced by the neurotransmitter,glutamate. Nonaka et al., PNAS 95: 2642-2647 (1998). Glutamate-inducedneuronal excitotoxicity is also believed to be a major cause ofneurodegeneration associated with acute damage, such as in cerebralischemia, traumatic brain injury and bacterial infection. Furthermore itis believed that excessive glutamate signaling is a factor in thechronic neuronal damage seen in diseases such as Alzheimer's,Huntingdon's, Parkinson's, AIDS associated dementia, amyotrophic lateralsclerosis (AML) and multiple sclerosis (MS). Thomas, J. Am. Geriatr.Soc. 43: 1279-89 (1995). Consequently GSK3 inhibitors are believed to bea useful treatment in these and other neurodegenerative disorders.

Immune Potentiation

GSK3 phosphorylates transcription factor NF-AT and promotes its exportfrom the nucleus, in opposition to the effect of calcineurin. Beals etal., Science 275:1930-33 (1997). Thus, GSK3 blocks early immune responsegene activation via NF-AT, and GSK3 inhibitors may tend to permit orprolong activation of immune responses. Thus GSK3 inhibitors arebelieved to prolong and potentiate the immunostimulatory effects ofcertain cytokines, and such an effect may enhance the potential of thosecytokines for tumor immunotherapy or indeed for immunotherapy ingeneral.

Other Disorders

Lithium also has other biological effects. It is a potent stimulator ofhematopoiesis, both in vitro and in vivo. Hammond et al., Blood 55:26-28 (1980). In dogs, lithium carbonate eliminated recurrentneutropenia and normalized other blood cell counts. Doukas et al. ExpHematol 14: 215-221 (1986). If these effects of lithium are mediatedthrough the inhibition of GSK3, GSK3 inhibitors may have even broaderapplications.

Since inhibitors of GSK3 are useful in the treatment of many diseases,the identification of new inhibitors of GSK3 would be highly desirable.

SUMMARY OF THE INVENTION

It has now been surprisingly discovered that glycogen synthase kinase 3(GSK3) activity can be inhibited in vitro or in vivo by certainpyrimidine and pyridine based derivatives. Accordingly, the presentinvention provides new compounds, compositions and methods of inhibitingthe activity of GSK3 in vitro and of treatment of GSK3 mediateddisorders in vivo. In one aspect, the present invention provides newcompounds having GSK3 inhibition activity of the following formula (I):

wherein:

W is optionally substituted carbon or nitrogen;

X and Y are independently selected from the group consisting ofnitrogen, oxygen, and optionally substituted carbon;

A is optionally substituted aryl or heteroaryl;

R₁, R′₁, R₂, R′₂, R₃, R′₃, R₄ and R′₄ are independently selected fromthe group consisting of hydrogen, hydroxyl, and optionally substitutedloweralkyl, cycloloweralkyl, cyclicaminoalkyl, alkylaminoalkyl,loweralkoxy, amino, alkylamino, alkylcarbonyl, arylcarbonyl,aralkylcarbonyl, heteroarylcarbonyl, heteroaralkylcarbonyl, aryl andheteroaryl, and R′₁, R′₂, R′₃ and R′₄ are independently selected fromthe group consisting of hydrogen, and optionally substituted loweralkyl;

R₅ and R₇ are independently selected from the group consisting ofhydrogen, halo, and optionally substituted loweralkyl, cycloalkyl,alkoxy, amino, aminoalkoxy,alkylcarbonylamino, arylcarbonylamino,aralkylcarbonylamino, heteroarylcarbonylamino,heteroaralkylcarbonylamino, cycloimido, heterocycloimido, amidino,cycloamidino, heterocycloamidino, guanidinyl, aryl, biaryl, heteroaryl,heterobiaryl, heterocycloalkyl, and arylsulfonamido;

R₆ is selected from the group consisting of hydrogen, hydroxy, halo,carboxyl, nitro, amino, amido, amidino, imido, cyano, and substituted orunsubstituted loweralkyl, loweralkoxy, alkylcarbonyl, arylcarbonyl,aralkylcarbonyl, heteroarylcarbonyl,beteroaralkylcarbonyl,alkylcarbonyloxy, arylcarbonyloxy,aralkylcarbonyloxy, beteroarylcarbonyloxy, heteroaralkylcarbonyloxy,alkylaminocarbonyloxy, arylaminocarbonyloxy, formyl, loweralkylcarbonyl,loweralkoxycarbonyl, aminocarbonyl, aminoaryl, alkylsulfonyl,sulfonamido, aminoalkoxy, alkylamino, heteroarylamino,alkylcarbonylamino, alkylaminocarbonylamino, arylaminocarbonylamino,aralkylcarbonylamino, heteroarylcarbonylamino, arylcarbonylamino,heteroarylcarbonylamino cycloamido, cyclothioamido, cycloamidino,heterocycloamidino, cycloimido, heterocycloimido, guanidinyl, aryl,heteroaryl, heterocyclo, heterocycloalkyl, arylsulfonyl andarylsulfonamido;

and the pharmaceutically acceptable salts thereof.

Presently particularly preferred and novel compounds of the inventionare provided by the compounds of formulas (IV) and (V):

wherein X, R₁-R₆, and R₈-R₁₄ have the meanings described above, and R₁₅is selected from the group consisting of hydrogen, nitro, cyano, amino,alkyl, halo, haloloweralkyl, alkyloxycarbonyl, aminocarbonyl,alkylsulfonyl and arylsulfonyl, and the pharmaceutically acceptablesalts thereof.

The methods, compounds and compositions of the invention may be employedalone, or in combination with other pharmacologically active agents inthe treatment of disorders mediated by GSK3 activity, such as in thetreatment of diabetes, Alzheimer's disease and other neurodegenerativedisorders, obesity, atherosclerotic cardiovascular disease, essentialhypertension, polycystic ovary syndrome, syndrome X, ischemia,especially cerebral ischemia, traumatic brain injury, bipolar disorder,immunodeficiency or cancer.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT

In accordance with the present invention, compounds, compositions andmethods are provided for the inhibition of glycogen synthase kinase 3(GSK3) activity, either in vitro or in vivo. In one aspect, the presentinvention provides new compounds having GSK3 inhibition activity of thefollowing formula (I):

wherein:

W is optionally substituted carbon or nitrogen;

X and Y are independently selected from the group consisting ofnitrogen, oxygen, and optionally substituted carbon;

A is optionally substituted aryl or heteroaryl;

R₁, R′₁, R₂, R′₂, R₃, R′₃, R₄ and R′₄ are independently selected fromthe group consisting of hydrogen, hydroxyl, and optionally substitutedloweralkyl, cycloloweralkyl, cyclicaminoalkyl, alkylaminoalkyl,loweralkoxy, amino, alkylamino, alkylcarbonyl, arylcarbonyl,aralkylcarbonyl, heteroarylcarbonyl, heteroaralkylcarbonyl, aryl andheteroaryl, and R′₁, R′₂, R′₃ and R′₄ are independently selected fromthe group consisting of hydrogen, and optionally substituted loweralkyl;

R₅ and R₇ are independently selected from the group consisting ofhydrogen, halo, and optionally substituted loweralkyl, cycloalkyl,alkoxy, amino, aminoalkoxy, alkylamino, aralkylamino,heteroaralkylamino, arylamino, heteroarylamino cycloimido,heterocycloimido, amidino, cycloamidino, heterocycloamidino, guanidinyl,aryl, biaryl, heteroaryl, heterobiaryl, heterocycloalkyl, andarylsulfonamido;

R₆ is selected from the group consisting of hydrogen, hydroxy, halo,carboxyl, nitro, amino, amido, amidino, imido, cyano, and substituted orunsubstituted loweralkyl, loweralkoxy, alkylcarbonyl, arylcarbonyl,aralkylcarbonyl, heteroarylcarbonyl, heteraralkylcarbonyl,alkylcarbonyloxy, arylcarbonyloxy, aralkylcarbonyloxy,alkylaminocarbonyloxy, arylaminocarbonyloxy, formyl, loweralkylcarbonyl,loweralkoxycarbonyl, aminocarbonyl, aminoaryl, alkylsulfonyl,sulfonamido, aminoalkoxy, alkylamino, heteroarylamino,alkylcarbonylamino, alkylaminocarbonylamino, arylaminocarbonylamino,aralkylcarbonylamino, heteroaralkylcarbonylamino, arylcarbonylamino,heteroarylcarbonylamino cycloamido, cyclothioamido, cycloamidino,heterocycloamidino, cycloimido, heterocycloimido, guanidinyl, aryl,heteroaryl, heterocyclo, heterocycloalkyl, arylsulfonyl andarylsulfonamido;

and the pharmaceutically acceptable salts thereof.

In one presently preferred embodiment of the invention, at least one ofX and Y is nitrogen. Representative compounds of this group includethose compounds in which one of X and Y is nitrogen and the other of Xand Y is oxygen or optionally substituted carbon. Preferably, both X andY are nitrogen.

The constituent A can be an aromatic ring having from 3 to 10 carbonring atoms and optionally 1 or more ring heteroatoms. Thus, in oneembodiment, A can be optionally substituted carbocyclic aryl.Alternatively, A is optionally substituted heteroaryl, such as, forexample, substituted or unsubstituted pyridyl, pyrimidinyl, thiazolyl,indolyl, imidazolyl, oxadiazolyl, tetrazolyl, pyrazinyl, triazolyl,thiophenyl, furanyl, quinolinyl, purinyl, naphthyl, benzothiazolyl,benzopyridyl, and benzimidazolyl, which may substituted with at leastone and not more than 3 substitution groups. Representative substitutiongroups can be independently selected from the group consisting of, forexample, nitro, amino, cyano, halo, thioamido, amidino, oxamidino,alkoxyamidino, imidino, guanidino, sulfonamido, carboxyl, formyl,loweralkyl, haloloweralkyl, loweralkoxy, haloloweralkoxy,loweralkoxyalkyl, loweralkylaminoloweralkoxy, loweralkylcarbonyl,loweraralkylcarbonyl, lowerheteroaralkylcarbonyl, alkylthio, aminoalkyland cyanoalkyl.

In a presently particularly preferred embodiment of the invention, A hasthe formula:

wherein R₈ and R₉ are independently selected from the group consistingof hydrogen, hydroxy, nitro, amino, cyano, halo, thioamido, amidino,oxamidino, alkoxyamidino, imidino, guanidinyl, sulfonamido, carboxyl,fomyl, loweralkyl, aminoloweralkyl, loweralkylaminoloweralkyl,haloloweralkyl, loweralkoxy, haloloweralkoxy, loweralkoxyalkyl,loweralkylaminoloweralkoxy, loweralkylcarbonyl, loweraralkylcarbonyl,lowerheteroaralkylcarbonyl, alkylthio, aryl and, aralkyl. Mostpreferably, A is selected from the group consisting of aminopyridyl,nitropyridyl, aminonitropyridyl, cyanopyridyl, cyanothiazolyl,aminocyanopyridyl, trifluoromethylpyridyl, methoxypyridyl,methoxynitropyridyl, methoxycyanopyridyl and nitrothiazolyl.

In other embodiments of the invention at least one of R₁, R′₁, R₂, R′₂,R₃, R′₃, R₄, and R′₄ may be hydrogen, or unsubstituted or substitutedloweralkyl selected from the group consisting of haloloweralkyl,heterocycloaminoalkyl, and loweralkylamino-loweralkyl; orloweralkylaminoloweralkyl. Presently preferred embodiments of theinvention include compounds wherein R₁, R′₁, R₂, R′₂, R₃, R′₃ and R₄ arehydrogen and R′₄is selected from the group consisting of hydrogen,methyl, ethyl, aminoethyl, dimethylaminoethyl, pyridylethyl,piperidinyl, pyrrolidinylethyl, piperazinylethyl and morpholinylethyl.

Other presently preferred compounds of the invention include compoundsof formula (I) wherein at least one of R₅ and R₇ is selected from thegroup consisting of substituted and unsubstituted aryl, heteroaryl andbiaryl. In presently preferred embodiments, at least one of R₅ and R₇ isa substituted or unsubstituted moiety of the formula:

wherein R₁₀, R₁₁, R₁₂, R₁₃, and R₁₄ are independently selected from thegroup consisting of hydrogen, nitro, amino, -cyano, halo, thioamido,carboxyl, hydroxy, and optionally substituted loweralkyl, loweralkoxy,loweralkoxyalkyl, haloloweralkyl, haloloweralkoxy, aminoalkyl,alkylamino, aminoalkylalkynyl, alkylaminoalkylalkynyl, alkylthio,alkylcarbonylamino, aralkylcarbonylamino, heteroaralkylcarbonylamino,arylcarbonylamino, heteroarylcarbonylamino aminocarbonyl,loweralkylaminocarbonyl, aminoaralkyl, loweralkylaminoalkyl, aryl,heteroaryl, cycloheteroalkyl, aralkyl, alkylcarbonyloxy,arylcarbonyloxy, aralkylcarbonyloxy, arylcarbonyloxyalkyl,alkylcarbonyloxyalkyl, heteroarylcarbonyloxyalkyl,aralkycarbonyloxyalkyl, and heteroaralkcarbonyloxyalkyl. Presentlyparticularly preferred compounds are obtained wherein R₁₀, R₁₁, R₁₃, andR₁₄ are hydrogen and R₁₂ is selected from the group consisting of halo,loweralkyl, hydroxy, loweralkoxy, haloloweralkyl, aminocarbonyl,alkylaminocarbonyl and cyano; R₁₁, R₁₃, and R₁₄ are hydrogen and R₁₀ andR₁₂ are independently selected from the group consisting of halo,loweralkyl, hydroxy, loweralkoxy, haloloweralkyl and cyano; R₁₀, R₁₁,R₁₃, and R₁₄ are hydrogen and R₁₂ is heteroaryl; R₁₀, R₁₁, R₁₃, and R₁₄are hydrogen and R₁₂ is a heterocycloalkyl; and wherein at least one ofR₁₀, R₁₁, R₁₂, R₁₃, and R₁₄ are halo and the remainder of R₁₀, R₁₁, R₁₂,R₁₃, and R₁₄ are hydrogen. Preferably, at least one of R₅ and R₇ isselected from the group consisting of dichlorophenyl, difluorophenyl,trifluoromethylphenyl, chlorofluorophenyl, bromochlorophenyl,ethylphenyl, methylchlorophenyl, imidazolylphenyl, cyanophenyl,morphlinophenyl and cyanochlorophenyl.

In representative embodiments of the invention, R₆ may be substitutedalkyl, such as, for example, aralkyl, hydroxyalkyl, aminoalkyl,aminoaralkyl, carbonylaminoalkyl, alkylcarbonylaminoalkyl,arylcarbonylaminoalkyl, aralkylcarbonylaminoalkyl, aminoalkoxyalkyl andarylaminoalkyl; substituted amino such as alkylamino,alkylcarbonylamino, alkoxycarbonylamino, arylalkylamino,arylcarbonylamino, alkylthiocarbonylamino, arylsulfonylamino,heteroarylamino alkylcarbonylamino, arylcarbonylamino,heteroarylcarbonylamino, aralkylcarbonylamino, andheteroaralkylcarbonylamino; or substituted carbonyl such asunsubstituted or substituted aminocarbonyl, alkyloxycarbonyl,aryloxycarbonyl, aralkyloxycarbonyl and alkylaminoalkyloxycarbonyl. Inother embodiments, R₆ may be selected from the group consisting ofamidino, guanidino, cycloimido, heterocycloimido, cycloamido,heterocycloamido, cyclothioamido and heterocycloloweralkyl. In yet otherembodiments, R₆ may be aryl or heteroaryl, such as, for example,substituted or unsubstituted pyridyl, pyrimidinyl, piperazinyl,thiazolyl, indolyl, imidazolyl, oxadiazolyl, tetrazolyl, pyrazinyl,triazolyl, thienyl, furanyl, quinolinyl, pyrrolyopyridyl,benzothiazolyl, benzopyridyl, benzotriazolyl, and benzimidazolyl. In yetother embodiments, R₆ may be a monoketopiperazinyl group having thestructure:

wherein R₁₅ and R₁₆ are independently selected from the group consistingof hydrogen, loweralkyl, loweralkynyl, aryl, heteroaryl, arylloweralkyl,loweralkylarylloweralkyl, haloloweralkyl, haloarylloweralkyl carbocyclicand heterocyclic; or R₈ can be taken with another R₁₆ or with R₁₅ toform a carbocyclic, heterocyclic or aryl ring; and o is an integerbetween 1 and 6. In representative embodiments of this aspect of theinvention, R₁₅ is loweralkyl, such as methyl, ethyl, n-propyl,isopropyl, cyclopropyl, n-butyl, iso-butyl or t-butyl, or R₁₅ is takenwith R₁₆ to form a group having the structure:

Presently preferred, representative compounds of this group include, forexample,1-[2-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-4-(2,4-dichlorophenyl)-5-pyrimidinyl]-2-piperazinone,1-[2-{[2-({6-amino-5-[-hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-4-(2,4-dichlorophenyl)-5-pyrimidinyl]-4-ethyl-3-methyl-2-piperazinone,1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-2-(2,4-dichlorophenyl)-3-pyridinyl]-4-methyl-2-piperazinone,1-[2-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-4-(2,4-dichlorophenyl)-5-pyrimidinyl]-4-methyl-2-piperazinone,6-[(2-{[6-(2,4-dichlorophenyl)-5-(4-methyl-2-oxo-1-piperazinyl)-2-pyridinyl]amino}ethyl)amino]nicotinonitrile,1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-2-(4-ethylphenyl)-3-pyridinyl]-4-methyl-2-piperazinone,6-[(2-{[6-(4-ethylphenyl)-5-(4-methyl-2-oxo-1-piperazinyl)-2-pyridinyl]amino}ethyl)amino]nicotinonitrile,6-({2-[[6-(4-ethylphenyl)-5-(4-methyl-2-oxo-1-piperazinyl)-2-pyridinyl](methyl)amino]ethyl}amino)nicotinonitrile,6-[(2-{[4-(2,4-dichlorophenyl)-5-(4-methyl-2-oxo-1-piperazinyl)-2-pyrimidinyl]amino}ethyl)amino]nicotinonitrile,1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-2-(2,4-difluorophenyl)-3-pyridinyl]-4-methyl-2-piperazinone,4-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}arnino)ethyl]amino}-3-(4-methyl-2-oxo-1-piperazinyl)-2-pyridinyl]benzonitrile,1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-2-(2,4-dichlorophenyl)-3-pyridinyl]-4-isopropyl-2-piperazinone,1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-2-(2,4-dichlorophenyl)-3-pyridinyl]-4-ethyl-2-piperazinone,1-{6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-2-[2-(trifluoromethyl)phenyl]-3-pyridinyl}-4-methyl-2-piperazinone,1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-2-(2-bromophenyl)-3-pyridinyl]-4-methyl-2-piperazinone,1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)propyl]amino}-2-(2,4-dichlorophenyl)-3-pyridinyl]-4-methyl-2-piperazinone,1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)propyl]amino}-2-(2,4-dichlorophenyl)-3-pyridinyl]-4-methyl-2-piperazinone,1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)-1-methylethyl]amino}-2-(2,4-dichlorophenyl)-3-pyridinyl]-4-methyl-2-piperazinone,1-[6-{[2-({6-amino-5-[hydroxy(oxido)aminol-2-pyridinyl}amino)-1-methylethyl]amino}-2-(2,4-dichlorophenyl)-3-pyridinyl]-4-methyl-2-piperazinone,1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)-1,1-dimethylethyl]amino}-2-(2,4-dichlorophenyl)-3-pyridinyl]-4-methyl-2-piperazinone,1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl][2-(1-pyrrolidinyl)ethyl]amino}-2-(2,4-dichlorophenyl)-3-pyridinyl]-4-methyl-2-piperazinone,1-[6-[3-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)propyl]-2-(2,4-dichlorophenyl)-3-pyridinyl]-4-methyl-2-piperazinone,1-[6-[[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl](methyl)amino]-2-(2,4-dichlorophenyl)-3-pyridinyl]-4-methyl-2-piperazinone,1-[6-[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethoxy]-2-(2,4-dichlorophenyl)-3-pyridinyl]4-methyl-2-piperazinone,1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}oxy)ethyl]amino}-2-(2,4-dichlorophenyl)-3-pyridinyl]-4-methyl-2-piperazinone,1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl][2-(4-morpholinyl)ethyl]amino}-2-(2,4-dichlorophenyl)-3-pyridinyl]-4-methyl-2-piperazinone,1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-2-(2,4-dichlorophenyl)-3-pyridinyl]-2-piperazinone,1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-2-(2,4-dichlorophenyl)-3-pyridinyl]-4-cyclopropyl-2-piperazinone,1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-2-(2,4-dichlorophenyl)-3-pyridinyl]-4-cyclohexyl-2-piperazinone,1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-2-(4-bromophenyl)-3-pyridinyl]-4-methyl-2-piperazinone,1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-2-(2,4-dichlorophenyl)-3-pyridinyl]-3,4-dimethyl-2-piperazinone,1-(2-(2,4-dichlorophenyl)-6-{[2-({5-[hydroxy(oxido)amino]-6-methoxy-2-pyridinyl}amino)ethyl]amino}-3-pyridinyl)-4-methyl-2-piperazinone,1-(2-(2,4-dichlorophenyl)-6-{[2-({5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-3-pyridinyl)-4-methyl-2-piperazinone,4-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-3-(4-methyl-2-oxo-1-piperazinyl)-2-pyridinyl]benzamide,2-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-2-(2,4-dichlorophenyl)-3-pyridinyl]hexahydropyrrolo[1,2-a]pyrazin-3(4H)-one,1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-2-(2,4-dichlorophenyl)-3-pyridinyl]-4-(methylsulfonyl)-2-piperazinone,4-acetyl-1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-2-(2,4-dichlorophenyl)-3-pyridinyl]-2-piperazinone,2-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-2-(2,4-dichlorophenyl)-3-pyridinyl]hexahydropyrrolo[1,2-a]pyrazin-3(4H)-one,2-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-2-(2,4-dichlorophenyl)-3-pyridinyl]hexahydropyrrolo[1,2-a]pyrazin-3(4H)-one,2-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-2-(2,4-dichlorophenyl)-3-pyridinyl]hexahydropyrrolo[1,2-a]pyrazin-1(2H)-one,2-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-2-(2,4-dichlorophenyl)-3-pyridinyl]hexahydropyrrolo-[1,2-a]pyrazin-1(2H)-one,1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-2-(2,4-difluorophenyl)-3-pyridinyl]-4-ethyl-2-piperazinone,4-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-3-(4-ethyl-2-oxo-1-piperazinyl)-2-pyridinyl]benzonitrile,1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)propyl]amino}-2-(2,4-dichlorophenyl)-3-pyridinyl]-4-ethyl-2-piperazinone,1-(6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-2-phenyl-3-pyridinyl)-4-methyl-2-piperazinone,1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-2-(4-chlorophenyl)-3-pyridinyl]-4-methyl-2-piperazinone,3-amino-1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-2-(2,4-dichlorophenyl)-3-pyridinyl]-2-piperidinone,1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-2-(3-chlorophenyl)-3-pyridinyl]-4-methyl-2-piperazinone,1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-2-(2,4-dichlorophenoxy)-3-pyridinyl]-4-methyl-2-piperazinone,1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-2-(2,4-dichlorophenyl)-3-pyridinyl]-4-(2,2,2-trifluoroethyl)-2-piperazinone,4-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-2-(2,4-dichlorophenyl)-3-pyridinyl]-3-morpholinone,1-{6-[2-(6-Amino-5-nitro-pyridin-2-ylamino)ethylamino]-[2,3′]bipyridinyl-3-yl}-4-methyl-piperazin-2-one,1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-2-(2,4-dichlorophenyl)-3-pyridinyl]-2-piperidinone,1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-2-(4-chloro-2-methylphenyl)-3-pyridinyl]-4-methyl-2-piperazinone,1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-2-(4-methoxyphenyl)-3-pyridinyl]-4-methyl-2-piperazinone,and1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-2-(3-furyl)-3-pyridinyl]-4-methyl-2-piperazinone.

As used herein, representative heterocyclo groups include, for example,those shown below (where the point of attachment of the substituentgroup, and the other substituent groups shown below, is through theupper left-hand bond). These heterocyclo groups can be furthersubstituted and may be attached at various positions as will be apparentto those having skill in the organic and medicinal chemistry arts inconjunction with the disclosure herein.

Representative heteroaryl groups include, for example, those shownbelow. These heteroaryl groups can be further substituted and may beattached at various positions as will be apparent to those having skillin the organic and medicinal chemistry arts in conjunction with thedisclosure herein.

Representative cycloimido and heterocycloimido groups include, forexample, those shown below. These cycloimido and heterocycloimido can befurther substituted and may be attached at various positions as will beapparent to those having skill in the organic and medicinal chemistryarts in conjunction with the disclosure herein.

Representative substituted amidino and heterocycloamidino groupsinclude, for example, those shown below. These amidino andheterocycloamidino groups can be further substituted as will be apparentto those having skill in the organic and medicinal chemistry arts inconjunction with the disclosure herein.

Representative substituted alkylcarbonylamino, alkyloxycarbonylamino,aminoalkyloxycarbonylamino, and arylcarbonylamino groups include, forexample, those shown below. These groups can be further substituted aswill be apparent to those having skill in the organic and medicinalchemistry arts in conjunction with the disclosure herein.

Representative substituted aminocarbonyl groups include, for example,those shown below. These can heterocyclo groups be further substitutedas will be apparent to those having skill in the organic and medicinalchemistry arts in conjunction with the disclosure herein.

Representative substituted alkoxycarbonyl groups include, for example,those shown below. These alkoxycarbonyl groups can be furthersubstituted as will be apparent to those having skill in the organic andmedicinal chemistry arts in conjunction with the disclosure herein.

Presently particularly preferred compounds of the invention includecompounds having the structure:

wherein X, R₁-R₆, and R₈-R₁₄ have the meanings described above, and thepharmaceutically acceptable salts thereof. Presently preferred,representative compounds of this group include, for example,[4-(4-imidazolylphenyl)pyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine,4-[5-imidazolyl-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]benzenecarbonitrile,4-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-5-imidazolylpyrimidin-4-yl]benzenecarbonitrile,[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine,4-[2-({2-[(5-nitro-2-pyridyl)amino]ethyl}amino)-7a-hydro-1,2,4-triazolo[1,5-a]pyrimidin-7-yl]benzenecarbonitrile,{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl]amine,[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine,6-[(2-{[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carbonitrile,[5-benzotriazolyl-4-(2,4-dichlorophenyl)pyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine,[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]methan-1-ol,[4-(2,4-dichlorophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-5-yl]methan-1-ol,2-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]isoindoline-1,3-dione,[5-amino-4-(2,4-dichlorophenyl)pyrimidin-2-yl]{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amine,{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-morpholin-4-ylpyrimidin-2-yl]amine,{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}{4-(2,4-dichlorophenyl)-5-[5-(trifluoromethyl)(1,2,3,4-tetraazolyl)]pyrimidin-2-yl}amine,1-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]pyrrolidine-2,5-dione,[4-(2,4-dichlorophenyl)-5-pyrazolylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine,[4-(2,4-dichlorophenyl)-5-(4-methylimidazolyl)pyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine,[4-(2,4-dichlorophenyl)-5-(2,4-dimethylimidazolyl)pyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine,6-[(2-{[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carbonitrile,{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-(morpholin-4-ylmethyl)pyrimidin-2-yl]amine,{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-piperazinylpyrimidin-2-yl]amine,{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(4-ethylphenyl)-5-imidazolylpyrimidin-2-yl]amine,1-[4-(2,4-dichlorophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-5-yl]hydropyridin-2-one,[5-benzimidazolyl-4-(2,4-dichlorophenyl)pyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine,{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl]methylamine,{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-(4-pyridyl)pyrimidin-2-yl]amine,{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-(4-methylpiperazinyl)pyrimidin-2-yl]amine,[4-(2,4-dichlorophenyl)-5-(2-methylimidazolyl)pyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine,{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-(2-methylimidazolyl)pyrimidin-2-yl]amine,{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-(4-phenylimidazolyl)pyrimidin-2-yl]amine,{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-(2,4-dimethylimidazolyl)pyrimidin-2-yl]amine,[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl](2-{[5-(trifluoromethyl)(2-pyridyl)]amino}ethyl)amine,[4-(2,4-dichlorophenyl)-5-piperazinylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine,[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl][2-(dimethylamino)ethyl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine,1-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]-4-methylpiperazine-2,6-dione,[4-(2,4-dichlorophenyl)-5-(1-methylimidazol-2-yl)pyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine,1-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]-3-morpholin-4-ylpyrrolidine-2,5-dione,1-[4-(2,4-dichlorophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-5-yl]-4-methylpiperazine-2,6-dione,1-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]-3-(dimethylamino)pyrrolidine-2,5-dione,{5-imidazol-2-yl-4-[4-(trifluoromethyl)phenyl]pyrimidin-2-yl}{2-[(5-nitro(2-pyridyl))amino]ethyl}amine,{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-(1-methylimidazol-2-yl)pyrimidin-2-yl]amine,[4-(2,4-dichlorophenyl)-5-(4-methylpiperazinyl)pyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine,[4-(2,4-dichlorophenyl)-5-(morpholin-4-ylmethyl)pyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine,[4-(2,4-dichlorophenyl)-5-(4-methylimidazol-2-yl)pyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine,{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-(4-methylimidazol-2-yl)pyrimidin-2-yl]amine,{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2-chlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amine,[4-(2-chloro-4-fluorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine,[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}(2-pyrrolidinylethyl)amine,[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl](2-morpholin-4-ylethyl){2-[(5-nitro(2-pyridyl))amino]ethyl}amine,6-[(2-{[4-(2,4-dichlorophenyl)-5-(4-methylimidazol-2-yl)pyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carbonitrile,{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2-chloro-4-fluorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amine,[4-(4-ethylphenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine,[5-((1E)-1-aza-2-morpholin-4-ylprop-1-enyl)-4-(2,4-dichlorophenyl)pyrimidin-2-yl]{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amine,N-[4-(2,4-dichlorophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-5-yl]acetamide,[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(6-methoxy-5-nitro(2-pyridyl))amino]ethyl}amine,6-[(2-{[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl]methylamino}ethyl)amino]pyridine-3-carbonitrile,6-[(2-{[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]methylamino}ethyl)amino]pyridine-3-carbonitrile,[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]methyl{2-[(5-nitro(2-pyridyl))amino]ethyl}amine,6-[(2-{[4-(2-chloro-4-fluorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carbonitrile,[4-(4-chlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine,{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(4-chloro-2-methylphenyl)-5-imidazol-2-ylpyrimidin-2-yl]amine,{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(4-bromo-2-chlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amine,6-[(2-{[4-(4-bromo-2-chlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carbonitrile,6-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]-3-pyrrolino[3,4-b]pyridine-5,7-dione,N-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]-2-(methylamino)acetamide,{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(4-bromo-2-chlorophenyl)-5-(4-methylimidazol-2-yl)pyrimidin-2-yl]amine,6-[(2-{[4-(4-bromo-2-chlorophenyl)-5-(4-methylimidazol-2-yl)pyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carbonitrile,{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2-chloro-4-fluorophenyl)-5-(4-methylimidazol-2-yl)pyrimidin-2-yl]amine,and6-[(2-{[4-(2,4-dichlorophenyl)-5-(5-chloro-2-oxohydropyridyl)pyrimidin-2-yl]aminoethyl)amino]pyridine-3-carbonitrile.

Other presently particularly preferred compounds of the inventioninclude compounds having the structure:

wherein X, R₁-R₆, and R₈-R₁₄ have the meanings described above, and R₁₅is selected from the group consisting of hydrogen, nitro, cyano, amino,alkyl, halo, haloloweralkyl, alkyloxycarbonyl, aminocarbonyl,alkylsulfonyl and arylsulfonyl, and the pharmaceutically acceptablesalts thereof. Presently preferred, representative compounds of thisgroup include, for example,[6-(2,4-dichlorophenyl)-5-imidazolyl(2-pyridyl)]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine,{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[6-(2,4-dichlorophenyl)-5-imidazolyl(2-pyridyl)]amine,6-[(2-{[6-(2,4-dichlorophenyl)-5-imidazolyl-2-pyridyl]amino}ethyl)amino]pyridine-3-carbonitrile,{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[6-(2,4-dichlorophenyl)-5-nitro(2-pyridyl)]amine,{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[6-(2,4-dichlorophenyl)-5-(4-methylimidazolyl)(2-pyridyl)]amine,6-[(2-{[6-(2,4-dichlorophenyl)-5-(4-methylimidazolyl)-2-pyridyl]amino}ethyl)amino]pyridine-3-carbonitrile,and[4-(4-bromo-2-chlorophenyl)-5-imidazol-2-yl-pyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine.

In another aspect, the invention provides compositions comprising anamount of a compound of formula (I) effective to modulate GSK3 activityin a human or animal subject when administered thereto, together with apharmaceutically acceptable carrier.

In yet other embodiments, the invention provides methods of inhibitingGSK3 activity in a human or animal subject, comprising administering tothe human or animal subject a GSK3 inhibitory amount of a compound ofstructure (I).

The present invention further provides methods of treating human oranimal subjects suffering from GSK3-mediated disorder in a human oranimal subject, comprising administering to the human or animal subjecta therapeutically effective amount of a compound of formula (I) above,either alone or in combination with other therapeutically active agents.

In yet other embodiments, the present invention provides compounds offormulas I, IV and V, as described above, for use as a pharmaceutical,as well as methods of use of those compounds in the manufacture of amedicament for the treatment of diabetes, Alzheimer's disease and otherneurodegenerative disorders, obesity, atherosclerotic cardiovasculardisease, essential hypertension, polycystic ovary syndrome, syndrome X,ischemia, especially cerebral ischemia, traumatic brain injury, bipolardisorder, immunodeficiency or cancer.

As used above and elsewhere herein the following terms have the meaningsdefined below:

“Glycogen synthase kinase 3” and “GSK3” are used interchangeably hereinto refer to any protein having more than 60% sequence homology to theamino acids between positions 56 and 340 of the human GSK3 beta aminoacid sequence (Genbank Accession No. L33801). To determine the percenthomology of two amino acid sequences or of two nucleic acids, thesequences are aligned for optimal comparison purposes (e.g., gaps can beintroduced in the sequence of one polypeptide or nucleic acid foroptimal alignment with the other polypeptide or nucleic acid). The aminoacid residues or nucleotides at corresponding amino acid positions ornucleotide positions are then compared. When a position in one sequenceis occupied by the same amino acid residue or nucleotide as thecorresponding position in the other sequence, then the molecules arehomologous at that position (i.e., as used herein amino acid or nucleicacid “homology” is equivalent to amino acid or nucleic acid “identity”).The percent homology between the two sequences is a function of thenumber of identical positions shared by the sequences (i.e., %homology=# of identical positions/total # of positions×100). GSK3 wasoriginally identified by its phosphorylation of glycogen synthase asdescribed in Woodgett et al., Trends Biochem. Sci., 16:177-81 (1991),incorporated herein by reference. By inhibiting GSK3 kinase activity,activities downstream of GSK3 activity may be inhibited, or,alternatively, stimulated. For example, when GSK3 activity is inhibited,glycogen synthase may be activated, resulting in increased glycogenproduction. GSK3 is also known to act as a kinase in a variety of othercontexts, including, for example, phosphorylation of c-jun, β-catenin,and tau protein. It is understood that inhibition of GSK3 kinaseactivity can lead to a variety of effects in a variety of biologicalcontexts. The invention, however, is not limited by any theories ofmechanism as to how the invention works.

“GSK3 inhibitor” is used herein to refer to a compound that exhibits anIC₅₀ with respect to GSK3 of no more than about 100 μM and moretypically not more than about 50 μM, as measured in the cell-free assayfor GSK3 inhibitory activity described generally hereinbelow. “IC₅₀” isthat concentration of inhibitor which reduces the activity of an enzyme(e.g., GSK3) to half-maximal level. Representative compounds of thepresent invention have been discovered to exhibit inhibitory activityagainst GSK3. Compounds of the present invention preferably exhibit anIC₅₀ with respect to GSK3 of no more than about 10 μM, more preferably,no more than about 5 μM, even more preferably not more than about 1 μM,and most preferably, not more than about 200 nM, as measured in thecell-free GSK3 kinase assay.

“Optionally substituted” refers to the replacement of hydrogen with amonovalent or divalent radical. Suitable substitution groups include,for example, hydroxyl, nitro, amino, imino, cyano, halo, thio,thioamido, amidino, imidino, oxo, oxamidino, methoxamidino, imidino,guanidino, sulfonamido, carboxyl, formyl, loweralkyl, haloloweralkyl,loweralkoxy, haloloweralkoxy, loweralkoxyalkyl,alkylcarbonyl,arylcarbonyl, aralkylcarbonyl, heteroarylcarbonyl,heteroaralkylcarbonyl, alkylthio, aminoalkyl, cyanoalkyl, and the like.

The substitution group can itself be substituted. The group substitutedonto the substitution group can be carboxyl, halo; nitro, amino, cyano,hydroxyl, loweralkyl, loweralkoxy, aminocarbonyl, —SR, thioamido, —SO₃H,—SO₂R or cycloalkyl, where R is typically hydrogen, hydroxyl orloweralkyl.

When the substituted substituent includes a straight chain group, thesubstitution can occur either within the chain (e.g., 2-hydroxypropyl,2-aminobutyl, and the like) or at the chain terminus (e.g.,2-hydroxyethyl, 3-cyanopropyl, and the like). Substituted substitutentscan be straight chain, branched or cyclic arrangements of covalentlybonded carbon or heteroatoms.

“Loweralkyl” as used herein refers to branched or straight chain alkylgroups comprising one to ten carbon atoms that are unsubstituted orsubstituted, e.g., with one or more halogen, hydroxyl or other groups,including, e.g., methyl, ethyl, propyl, isopropyl, n-butyl, t-butyl,neopentyl, trifluoromethyl, pentafluoroethyl and the like.

“Alkylenyl” refers to a divalent straight chain or branched chainsaturated aliphatic radical having from 1 to 20 carbon atoms. Typicalalkylenyl groups employed in compounds of the present invention areloweralkylenyl groups that have from 1 to about 6 carbon atoms in theirbackbone. “Alkenyl” refers herein to straight chain, branched, or cyclicradicals having one or more double bonds and from 2 to 20 carbon atoms.“Alkynyl” refers herein to straight chain, branched, or cyclic radicalshaving one or more triple bonds and from 2 to 20 carbon atoms.

“Loweralkoxy” as used herein refers to RO— wherein R is loweralkyl.Representative examples of loweralkoxy groups include methoxy, ethoxy,t-butoxy, trifluoromethoxy and the like.

“Cycloalkyl” refers to a mono- or polycyclic, heterocyclic orcarbocyclic alkyl substituent. Typical cycloalkyl substituents have from3 to 8 backbone (i.e., ring) atoms in which each backbone atom is eithercarbon or a heteroatom. The term “heterocycloalkyl” refers herein tocycloalkyl substituents that have from 1 to 5, and more typically from 1to 4 heteroatoms in the ring structure. Suitable heteroatoms employed incompounds of the present invention are nitrogen, oxygen, and sulfur.Representative heterocycloalkyl moieties include, for example,morpholino, piperazinyl, piperadinyl and the like. Carbocycloalkylgroups are cycloalkyl groups in which all ring atoms are carbon. Whenused in connection with cycloalkyl substituents, the term “polycyclic”refers herein to fused and non-fused alkyl cyclic structures.

“Halo” refers herein to a halogen radical, such as fluorine, chlorine,bromine or iodine. “Haloalkyl” refers to an alkyl radical substitutedwith one or more halogen atoms. The term “haloloweralkyl” refers to aloweralkyl radical substituted with one or more halogen atoms. The term“haloalkoxy” refers to an alkoxy radical substituted with one or morehalogen atoms. The term “haloloweralkoxy” refers to a loweralkoxyradical substituted with one or more halogen atoms.

“Aryl” refers to monocyclic and polycyclic aromatic groups having from 3to 14 backbone carbon or hetero atoms, and includes both carbocyclicaryl groups and heterocyclic aryl groups. Carbocyclic aryl groups arearyl groups in which all ring atoms in the aromatic ring are carbon. Theterm “heteroaryl” refers herein to aryl groups having from 1 to 4heteroatoms as ring atoms in an aromatic ring with the remainder of thering atoms being carbon atoms. When used in connection with arylsubstituents, the term “polycyclic” refers herein to fused and non-fusedcyclic structures in which at least one cyclic structure is aromatic,such as, for example, benzodioxozolo (which has a heterocyclic structurefused to a phenyl group, i.e.

naphthyl, and the like. Exemplary aryl moieties employed as substituentsin compounds of the present invention include phenyl, pyridyl,pyrimidinyl, thiazolyl, indolyl, imidazolyl, oxadiazolyl, tetrazolyl,pyrazinyl, triazolyl, thiophenyl, furanyl, quinolinyl, purinyl,naphthyl, benzothiazolyl, benzopyridyl, and benzimidazolyl, and thelike.

“Aralkyl” refers to an alkyl group substituted with an aryl group.Typically, aralkyl groups employed in compounds of the present inventionhave from 1 to 6 carbon atoms incorporated within the alkyl portion ofthe aralkyl group. Suitable aralkyl groups employed in compounds of thepresent invention include, for example, benzyl, picolyl, and the like.

“Amino” refers herein to the group —NH₂. The term “alkylamino” refersherein to the group —NRR′ where R and R′ are each independently selectedfrom hydrogen or a lower alkyl. The term “arylamino” refers herein tothe group —NRR′ where R is aryl and R′ is hydrogen, a lower alkyl, or anaryl. The term “aralkylamino” refers herein to the group —NRR′ where Ris a lower aralkyl and R′ is hydrogen, a loweralkyl, an aryl, or aloweraralkyl.

The term “arylcycloalkylamino” refers herein to the group,aryl-cycloalkyl-NH—, where cycloalkyl is a divalent cycloalkyl group.Typically, cycloalkyl has from 3 to 6 backbone atoms, of which,optionally 1 to about 4 are heteroatoms. The term “aminoalkyl” refers toan alkyl group that is terminally substituted with an amino group.

The term “alkoxyalkyl” refers to the group -alk₁-O-alk₂ where alk₁ isalkylenyl or alkenyl, and alk₂ is alkyl or alkenyl. The term“loweralkoxyalkyl” refers to an alkoxyalkyl where alk₁ is loweralkylenylor loweralkenyl, and alk₂ is loweralkyl or loweralkenyl. The term“aryloxyalkyl” refers to the group -alkylenyl-O-aryl. The term“aralkoxyalkyl” refers to the group -alkylenyl-O-aralkyl, where aralkylis a loweraralkyl.

The term “alkoxyalkylamino” refers herein to the group—NR—(alkoxylalkyl), where R is typically hydrogen, loweraralkyl, orloweralkyl. The term “aminoloweralkoxyalkyl” refers herein to anaminoalkoxyalkyl in which the alkoxyalkyl is a loweralkoxyalkyl.

The term “aminocarbonyl” refers herein to the group —C(O)—NH₂.“Substituted aminocarbonyl” refers herein to the group —C(O)—NRR′ whereR is loweralkyl and R′ is hydrogen or a loweralkyl. The term“arylaminocarbonyl” refers herein to the group —C(O)—NRR′ where R is anaryl and R′ is hydrogen, loweralkyl or aryl. “aralkylaminocarbonyl”refers herein to the group —C(O)—NRR′ where R is loweraralkyl and R′ ishydrogen, loweralkyl, aryl, or loweraralkyl.

“Aminosulfonyl” refers herein to the group —S(O)₂—NH₂. “Substitutedaminosulfonyl” refers herein to the group —S(O)₂—NRR′ where R isloweralkyl and R′ is hydrogen or a loweralkyl. The term“aralkylaminosulfonlyaryl” refers herein to the group-aryl-S(O)₂—NH-aralkyl, where the aralkyl is loweraralkyl.

“Carbonyl” refers to the divalent group —C(O)—.

“Carbonyloxy” refers generally to the group —C(O)—O—,. Such groupsinclude esters, —C(O)—O—R, where R is loweralkyl, cycloalkyl, aryl, orloweraralkyl. The term “carbonyloxycycloalkyl” refers generally hereinto both an “carbonyloxycarbocycloalkyl” and an“carbonyloxyheterocycloalkyl”, i.e., where R is a carbocycloalkyl orheterocycloalkyl, respectively. The term “arylcarbonyloxy” refers hereinto the group —C(O)—O-aryl, where aryl is a mono- or polycyclic,carbocycloaryl or heterocycloaryl. The term “aralkylcarbonyloxy” refersherein to the group —C(O)—O-aralkyl, where the aralkyl is loweraralkyl.

The term “sulfonyl” refers herein to the group —SO₂—. “Alkylsulfonyl”refers to a substituted sulfonyl of the structure —SO₂R—in which R isalkyl. Alkylsulfonyl groups employed in compounds of the presentinvention are typically loweralkylsulfonyl groups having from 1 to 6carbon atoms in its backbone structure. Thus, typical alkylsulfonylgroups employed in compounds of the present invention include, forexample, methylsulfonyl (i.e., where R is methyl), ethylsulfonyl (i.e.,where R is ethyl), propylsulfonyl (i.e., where R is propyl), and thelike. The term “arylsulfonyl” refers herein to the group —SO₂-aryl. Theterm “aralkylsulfonyl” refers herein to the group —SO₂-aralkyl, in whichthe aralkyl is loweraralkyl. The term “sulfonamido” refers herein to—SO₂NH₂.

As used herein, the term “carbonylamino” refers to the divalent group—NH—C(O)— in which the hydrogen atom of the amide nitrogen of thecarbonylamino group can be replaced a loweralkyl, aryl, or loweraralkylgroup. Such groups include moieties such as carbamate esters(—NH—C(O)—O—R) and amides —NH—C(O)—O—R, where R is a straight orbranched chain loweralkyl, cycloalkyl, or aryl or loweraralkyl. The term“loweralkylcarbonylamino” refers to alkylcarbonylamino where R is aloweralkyl having from 1 to about 6 carbon atoms in its backbonestructure. The term “arylcarbonylamino” refers to group —NH—C(O)—R whereR is an aryl. Similarly, the term “aralkylcarbonylamino” refers tocarbonylamino where R is a lower aralkyl.

As used herein, the term “guanidino” or “guanidyl” refers to moietiesderived from guanidine, H₂N—C(═NH)—NH₂. Such moieties include thosebonded at the nitrogen atom carrying the formal double bond (the“2”-position of the guanidine, e.g., diaminomethyleneamino, (H₂N)₂C═NH—)and those bonded at either of the nitrogen atoms carrying a formalsingle bond (the “1-” and/or “3”-positions of the guandine, e.g.,H₂N—C(═NH)—NH—). The hydrogen atoms at any of the nitrogens can bereplaced with a suitable substituent, such as loweralkyl, aryl, orloweraralkyl.

As used herein, the term “amidino” refers to the moieties R—C(═N)—NR′—(the radical being at the “N¹” nitrogen) and R(NR′)C═N— (the radicalbeing at the “N²” nitrogen), where R and R′ can be hydrogen, loweralkyl,aryl, or loweraralkyl.

Compounds of the present invention can be readily synthesized using themethods described herein, or other methods, which are well known in theart. For example, the synthesis of pyrimidines having a wide variety ofsubstituents is comprehensibly reviewed in D. J. Brown, “ThePyrimidines,” vol. 54, Wiley (1994), which is incorporated herein byreference. The compounds described herein were synthesized using bothsolution-phase and resin-based (i.e., solid-phase) techniques.

Pyrimidine based compounds of the present invention can be readilysynthesized in solution by reaction of a carbonyl-containing derivativewith N,N-dimethylforrnamide dimethyl acetal (DMFDMA). The intermediateenaminoketone that results is then reacted with a guanidine in thepresence of an organic solvent and a suitable base such as sodiumethoxide, sodium methoxide, sodium hydroxide or cesium carbonate atvarious temperatures to give a pyrimidine. This method is generallydescribed in Menozzi et al., J. Heterocyclic Chem., 24:1669 (1987), P.Schenone et al., J. Heterocyclic Chem., 27:295 (1990), R. Paul et al.,J. Med. Chem., 36: 2716 (1993) and J. Zimmermann et al., Arch. Pharm.,329:371 (1996), all of which are incorporated herein by reference.

Carbonyl-containing starting reagents that are suitable for use in thisreaction scheme include, for example, β-keto esters, alkyl aryl ketones,β-keto sulfones, α-nitro ketones, β-keto nitriles, desoxybenzoins, arylheteroarylmethyl ketones, and the like. The carbonyl-containing startingreagents can either be purchased or synthesized using known methods.

For example, β-keto esters can be readily synthesized by reaction of anacid chloride or other activated carboxylic acid with potassium ethylmalonate in the presence of triethylamine in accordance with the methoddescribed in R. J. Clay et al., Synthesis, 1992:290 (1992), which isincorporated herein by reference. Alternatively, the desired β-ketoester can be synthesized by deprotonating an appropriate methyl ketonewith a suitable base such as sodium hydride, followed by condensationwith diethylcarbonate in accordance with the method described in Sircaret al., J. Med. Chem., 28:1405 (1985), which is incorporated herein byreference.

Likewise, β-keto sulfones and a-nitro ketones can be prepared usingknown methods, such as those described in N. S. Simpkins, “Sulphones inOrganic Synthesis,” Pergamon (1993) (β-keto sulfones) and M. Jung et.al., J. Org. Chem., 52:4570 (1987) (α-nitro ketones), both of which areincorporated herein by reference. β-keto nitrites can be readilyprepared by reaction of an α-halo ketone with sodium or potassiumcyanide.

When the substrate is a doubly activated carbonyl compound (e.g., β-ketoester, β-keto sulfone, β-keto nitrile, and the like) the firstcondensation is typically conducted with a small excess of DMFDMA in asolvent such as THF at 70-80° C. for several hours. This method isdescribed in more detail in Example 25 hereinbelow (i.e., “SolutionMethod A”).

When a mono-activated substrate such as a methyl ketone is involved,DMFDMA is often used as the solvent at a higher temperature (90-100° C.)for a longer period of time (e.g., overnight). After completion of thecondensation reaction, the solvent and excess DMFDMA are removed invacuo. The resulting solid or oil is dissolved in an appropriate solventand heated with an equimolar amount of the guanidine and base. Thismethod is described in more detail in Example 60 hereinbelow (i.e.,“Solution Method B”).

When esters are formed, alkaline or acidic hydrolysis of the resultingpyrimidine yields the corresponding carboxylic acid. This acid can thenbe further coupled to various alcohols or amines to provide a variety ofester or amide derivatives.

Guanidines employed in the synthesis of invention compounds can bepurchased or, alternatively, synthesized by reacting the correspondingamine with a guanidino transfer reagent, such as, for example,benzotriazole carboxamidinium 4-methylbenzenesulfonate. This guanidinotransfer reagent is described in A. R. Katritzky et al., 1995, SyntheticCommunications, 25:1173 (1995), which is incorporated herein byreference. Thus, for example, benzotriazole carboxamidinium4-methylbenzenesulfonate can be reacted in equimolar quantity with anamine and one equivalent of diisopropyl ethyl amine (DIEA) inacetonitrile at room temperature overnight to yield guanidinium4-methylbenzenesulfonate upon addition of diethyl ether. Aminescontaining a nitrogen heterocyclic aryl can be prepared by nucleophilicsubstitution of a halo-substituted nitrogen heterocyclic aryl with anappropriate diamine, such as, for example, ethylenediamine orpropylenediamine. These diamines are particularly suitable for use asreaction solvents at reaction temperatures in the range of about 25° C.to 125° C. The preparation of specialized amines is noted in theExamples provided herein.

Other known synthesis methods can be used to prepare compounds of thepresent invention. For example, 5-aryl 2-aminopyrimidine can be preparedby reacting a guanidine with a vinamidinium salt, in accordance with themethod described in R. M. Wagner and C. Jutz, Chem. Berichte, p. 2975(1971), which is incorporated herein by reference. This method isillustrated in Example 67 hereinbelow (i.e., “Solution Method C”).

Similarly, 4-anilo-2-chloropyrimidine can be prepared by reactinganiline with 2,4-dichloropyrimidine. Likewise, an aniline can be treatedwith a 2,4-dichloropyrimidine to give the 4-anilo-2-chloropyrimidine.Further substitution with a second amine gives2-amino-4-aminopyrimidine.

In addition to solution-phase synthesis methods, solid-support(including resin-based) synthesis methods can also be used to synthesizecompounds of the present invention, especially for parallel andcombinatorial synthesis methodologies. For example, the synthesis oftetra-substituted pyrimidines may begin with the loading of an aromaticcarboxylic acid aldehyde, such as, for example, 4-formyl benzoic acid,to the amino group of a suitable resin, such as, for example, Rink amideresin (Novabiochem, San Diego, Calif.) (“Resin Method A” which isdescribed in more detail in Example 2). Knoevenagel condensation of aβ-keto ester gives an unsaturated intermediate that can be condensedwith 1H-pyrazole-1-carboxamidine hydrochloride (Aldrich) in the presenceof a suitable base (e.g., potassium carbonate). The intermediatedihydropyrimidine can then be oxidized to the resin bound pyrimidinewith 2,3-dichloro-5,6-dicyano-1,4-benzoquinone (DDQ) in benzene.Finally, substitution of the pyrazolo moiety by heating with an amine in1-methylpyrrolidone (NMP) or other suitable solvent is followed byacidolytic cleavage to give the desired pyrimidine. This synthesismethod can be used to generate pyrimidines with a substituent in the4-position of the pyrimidine ring.

Resin Method B, which is described in detail in Example 3, can be usedto synthesize pyrimidines in which the 6-position is unsubstituted. Ahydroxymethyl-resin, such as commercially available Sasrin resin (BachemBiosciences, King of Prussia, Pa.), is treated with triphenylphosphinedibromide in dichloromethane to convert the hydroxymethyl group on theresin to a bromomethyl group, as generally described in K. Ngu et al.,Tetrahedron Letters, 38:973 (1997), which is incorporated herein byreference. The bromine is then displaced by reaction with a primaryamine in NMP (at room temperature or 70-80° C.). The amine is thencoupled with the appropriate aromatic compound containing an acetylgroup. The coupling can be carried out with PyBOP® (Novabiochem, SanDiego, Calif.), and 4-methylmorpholine in NMP.

Resin Method B can also be used to incorporate an amino acid residueinto the resulting pyrimidine. For example, amino resin can be coupledto a 9-fluorenyl-methoxycarbonyl (FMOC)-protected amino acid usingstandard peptide synthesis conditions and methods. Further coupling with4-acetylbenzoic acid followed by reaction with N,N-dimethylformamidedimethyl acetal and cyclization with a guanidine produces a pyrimidinederivative having an amino acid residue incorporated within it.

Pyrimidines having e.g., a carboxamidophenyl group at position 6 andhydrogen at position 5 can be prepared from an amino (i.e.,—NH₂)-containing resin such as Rink amide resin (Novabiochem, San Diego,Calif.). This method is described in more detail in Example 10hereinbelow (“Resin Method C”).

Compounds of the present invention can also be prepared according toResin Method D, to produce 2,4-diaminopyrimidines. Resin-bound amine isreacted with a 2,4-dichloropyrimidine to give a resin-bound6-amino-2-chloropyrimidine. The resin-bound amine can be derived fromany suitable primary amine; however, anilines generally are notsuitable. Displacement with a second amine and cleavage of the productfrom the resin gives a 2,4-diaminopyrimidine. For the seconddisplacement, primary or secondary amines that may contain otherfunctional groups, such as unprotected hydroxy groups, are suitable. Theresulting dichloropyrimidine may be further substituted, for example,with an ester group at the 5-position. A 2,6-dichloropyridine can beused instead of 2,4-dichloropyrimidine to produce a 2,6-diaminopyridine.This scheme is described in more detail in Examples 17-19 hereinbelow.

Resin Method E can be used to produce a 2,6-diaminopyridine. The methodis analogous to Resin Method D except that a 2,6-dichloropyridine isused as the electrophile and the final product is a 2,6-diaminopyridine.Resin Method E is described in more detail in Examples 20-21hereinbelow.

Resin Method F can be used to synthesize 5-amino substituted compoundsof the present invention. Resin-bound amine is reacted with a halomethylaryl ketone. The resulting resin-bound aminomethyl ketone is thentreated with DMFDMA (neat) followed by cyclization with a guanidine togive the 2,5-diamino-6-arylpyrimidine. Resin Method F is described inmore detail in Example 22, hereinbelow.

Resin Method G, which is described in more detail in Example 23, can beused to synthesize compounds of the present invention having a carboxylgroup at the 5-position.

GSK3 inhibitor compounds of the present invention can be purified usingknown methods, such as, for example, chromatography, crystallization,and the like.

Compounds of the present invention preferably exhibit inhibitoryactivity that is relatively substantially selective with respect toGSK3, as compared to at least one other type of kinase. As used herein,the term “selective” refers to a relatively greater potency forinhibition against GSK3, as compared to at least one other type ofkinase. Preferably, GSK3 inhibitors of the present invention areselective with respect to GSK3, as compared to at least two other typesof kinases. Kinase activity assays for kinases other than GSK3 aregenerally known. See e.g., Havlicek et al., J. Med. Chem., 40:408-12(1997), incorporated herein by reference. GSK3 selectivity can bequantitated according to the following: GSK3selectivity=IC_(50 (other kinase))÷IC_(50 (GSK3)), where a GSK3inhibitor is selective for GSK3 whenIC_(50 (other kinase))>IC_(50 (GSK3)). Thus, an inhibitor that isselective for GSK3 exhibits a GSK3 selectivity of greater than 1-foldwith respect to inhibition of a kinase other than GSK3. As used herein,the term “other kinase” refers to a kinase other than GSK3. Suchselectivities are generally measured in the cell-free assay described inExample 265.

Typically, GSK3 inhibitors of the present invention exhibit aselectivity of at least about 2-fold (i.e.,IC_(50 (other kinase))÷IC_(50 (GSK3))) for GSK3, as compared to anotherkinase and more typically they exhibit a selectivity of at least about5-fold. Usually, GSK3 inhibitors of the present invention exhibit aselectivity for GSK3, as compared to at least one other kinase, of atleast about 10-fold, desirably at least about 100-fold, and morepreferably, at least about 1000-fold.

GSK3 inhibitory activity can be readily detected using the assaysdescribed herein, as well as assays generally known to those of ordinaryskill in the art. Exemplary methods for identifying specific inhibitorsof GSK3 include both cell-free and cell-based GSK3 kinase assays. Acell-free GSK3 kinase assay detects inhibitors that act by directinteraction with the polypeptide GSK3, while a cell-based GSK3 kinaseassay may identify inhibitors that function by direct interaction withGSK3 itself, or by other mechanisms, including, for example,interference with GSK3 expression or with post-translational processingrequired to produce mature active GSK3 or alteration of theintracellular localization of GSK3.

In general, a cell-free GSK3 kinase assay can be readily carried out by:(1) incubating GSK3 with a peptide substrate, radiolabeled ATP (such as,for example, γ³³P- or γ³²P-ATP, both available from Amersham, ArlingtonHeights, Ill.), magnesium ions, and optionally, one or more candidateinhibitors; (2) incubating the mixture for a period of time to allowincorporation of radiolabeled phosphate into the peptide substrate byGSK3 activity; (3) transferring all or a portion of the enzyme reactionmix to a separate vessel, typically a microtiter well that contains auniform amount of a capture ligand that is capable of binding to ananchor ligand on the peptide substrate; (4) washing to remove unreactedradiolabeled ATP; then (5) quantifying the amount of ³³P or ³²Premaining in each well. This amount represents the amount ofradiolabeled phosphate incorporated into the peptide substrate.Inhibition is observed as a reduction in the incorporation ofradiolabeled phosphate into the peptide substrate.

Suitable peptide substrates for use in the cell free assay may be anypeptide, polypeptide or synthetic peptide derivative that can bephosphorylated by GSK3 in the presence of an appropriate amount of ATP.Suitable peptide substrates may be based on portions of the sequences ofvarious natural protein substrates of GSK3, and may also containN-terminal or C-terminal modifications or extensions including spacersequences and anchor ligands. Thus, the peptide substrate may residewithin a larger polypeptide, or may be an isolated peptide designed forphosphorylation by GSK3.

For example, a peptide substrate can be designed based on a subsequenceof the DNA binding protein CREB, such as the SGSG-linked CREB peptidesequence within the CREB DNA binding protein described in Wang et al.,Anal. Biochem., 220:397-402 (1994), incorporated herein by reference. Inthe assay reported by Wang et al., the C-terminal serine in the SXXXSmotif of the CREB peptide is enzymatically prephosphorylated bycAMP-dependent protein kinase (PKA), a step which is required to renderthe N-terrninal serine in the motif phosphorylatable by GSK3. As analternative, a modified CREB peptide substrate can be employed which hasthe same SXXXS motif and which also contains an N-tertninal anchorligand, but which is synthesized with its C-terminal serineprephosphorylated (such a substrate is available commercially fromChiron Technologies PTY Ltd., Clayton, Australia). Phosphorylation ofthe second serine in the SXXXS motif during peptide synthesis eliminatesthe need to enzymatically phosphorylate that residue with PKA as aseparate step, and incorporation of an anchor ligand facilitates captureof the peptide substrate after its reaction with GSK3.

Generally, a peptide substrate used for a kinase activity assay maycontain one or more sites that are phosphorylatable by GSK3, and one ormore other sites that are phosphorylatable by other kinases, but not byGSK3. Thus, these other sites can be prephosphorylated in order tocreate a motif that is phosphorylatable by GSK3. The term“prephosphorylated” refers herein to the phosphorylation of a substratepeptide with non-radiolabeled phosphate prior to conducting a kinaseassay using that substrate peptide. Such prephosphorylation canconveniently be performed during synthesis of the peptide substrate.

The SGSG-linked CREB peptide can be linked to an anchor ligand, such asbiotin, where the serine near the C terminus between P and Y isprephosphorylated. As used herein, the term “anchor ligand” refers to aligand that can be attached to a peptide substrate to facilitate captureof the peptide substrate on a capture ligand, and which functions tohold the peptide substrate in place during wash steps, yet allowsremoval of unreacted radiolabeled ATP. An exemplary anchor ligand isbiotin. The term “capture ligand” refers herein to a molecule which canbind an anchor ligand with high affinity, and which is attached to asolid structure. Examples of bound capture ligands include, for example,avidin- or streptavidin-coated microtiter wells or agarose beads. Beadsbearing capture ligands can be further combined with a scintillant toprovide a means for detecting captured radiolabeled substrate peptide,or scintillant can be added to the captured peptide in a later step.

The captured radiolabeled peptide substrate can be quantitated in ascintillation counter using known methods. The signal detected in thescintillation counter will be proportional to GSK3 activity if theenzyme reaction has been run under conditions where only a limitedportion (e.g., less than 20%) of the peptide substrate isphosphorylated. If an inhibitor is present during the reaction, GSK3activity will be reduced, and a smaller quantity of radiolabeledphosphate will thus be incorporated into the peptide substrate. Hence, alower scintillation signal will be detected. Consequently, GSK3inhibitory activity will be detected as a reduction in scintillationsignal, as compared to that observed in a negative control where noinhibitor is present during the reaction. This assay is described inmore detail in Example 265 hereinbelow.

A cell-based GSK3 kinase activity assay typically utilizes a cell thatcan express both GSK3 and a GSK3 substrate, such as, for example, a celltransformed with genes encoding GSK3 and its substrate, includingregulatory control sequences for the expression of the genes. Incarrying out the cell-based assay, the cell capable of expressing thegenes is incubated in the presence of a compound of the presentinvention. The cell is lysed, and the proportion of the substrate in thephosphorylated form is determined, e.g., by observing its mobilityrelative to the unphosphorylated form on SDS PAGE or by determining theamount of substrate that is recognized by an antibody specific for thephosphorylated form of the substrate. The amount of phosphorylation ofthe substrate is an indication of the inhibitory activity of thecompound, i.e., inhibition is detected as a decrease in phosphorylationas compared to the assay conducted with no inhibitor present. GSK3inhibitory activity detected in a cell-based assay may be due, forexample, to inhibition of the expression of GSK3 or by inhibition of thekinase activity of GSK3.

Thus, cell-based assays can also be used to specifically assay foractivities that are implicated by GSK3 inhibition, such as, for example,inhibition of tau protein phosphorylation, potentiation of insulinsignaling, and the like. For example, to assess the capacity of a GSK3inhibitor to inhibit Alzheimer's-like phosphorylation ofmicrotubule-associated protein tau, cells may be co-transfected withhuman GSK3β and human tau protein, then incubated with one or morecandidate inhibitors. Various mammalian cell lines and expressionvectors can be used for this type of assay. For instance, COS cells maybe transfected with both a human GSK3β expression plasmid according tothe protocol described in Stambolic et al., 1996, Current Biology6:1664-68, which is incorporated herein by reference, and an expressionplasmid such as pSG5 that contains human tau protein coding sequenceunder an early SV40 promoter. See also Goedert et al., EMBO J.,8:393-399 (1989), which is incorporated herein by reference.Alzheimer's-like phosphorylation of tau can be readily detected with aspecific antibody such as, for example, AT8, which is available fromPolymedco Inc. (Cortlandt Manor, N.Y.) after lysing the cells. Thisassay is described in greater detail in the examples, hereinbelow.

Likewise, the ability of GSK3 inhibitor compounds to potentiate insulinsignaling by activating glycogen synthase can be readily ascertainedusing a cell-based glycogen synthase activity assay. This assay employscells that respond to insulin stimulation by increasing glycogensynthase activity, such as the CHO-HIRC cell line, which overexpresseswild-type insulin receptor (˜100,000 binding sites/cell). The CHO-HIRCcell line can be generated as described in Moller et al., J. Biol.Chem., 265:14979-14985 (1990) and Moller et al., Mol. Endocrinol.,4:1183-1191 (1990), both of which are incorporated herein by reference.The assay can be carried out by incubating serum-starved CHO-HIRC cellsin the presence of various concentrations of compounds of the presentinvention in the medium, followed by cell lysis at the end of theincubation period. Glycogen synthase activity can be detected in thelysate as described in Thomas et al., Anal Biochem., 25:486-499 (1968).Glycogen synthase activity is computed for each sample as a percentageof maximal glycogen synthase activity, as described in Thomas et al.,supra, and is plotted as a function of candidate GSK3 inhibitorconcentration. The concentration of candidate GSK3 inhibitor thatincreased glycogen synthase activity to half of its maximal level (i.e.,the EC₅₀) can be calculated by fitting a four parameter sigmoidal curveusing routine curve fitting methods that are well known to those havingordinary skill in the art. This is described in more detail in Example266, hereinbelow.

GSK3 inhibitors can be readily screened for in vivo activity such as,for example, using methods that are well known to those having ordinaryskill in the art. For example, candidate compounds having potentialtherapeutic activity in the treatment of type 2 diabetes can be readilyidentified by detecting a capacity to improve glucose tolerance inanimal models of type 2 diabetes. Specifically, the candidate compoundcan be dosed using any of several routes prior to administration of aglucose bolus in either diabetic mice (e.g. KK, db/db, ob/ob) ordiabetic rats (e.g. Zucker Fa/Fa or GK). Following administration of thecandidate compound and glucose, blood samples are removed at preselectedtime intervals and evaluated for serum glucose and insulin levels.Improved disposal of glucose in the absence of elevated secretion levelsof endogenous insulin can be considered as insulin sensitization and canbe indicative of compound efficacy. A detailed description of this assayis provided in the examples, hereinbelow.

The compounds of the present invention can be used in the form of saltsderived from inorganic or organic acids. These salts include but are notlimited to the following: acetate, adipate, alginate, citrate,aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, camphorate,camphorsulfonate, digluconate, cyclopentanepropionate, dodecylsulfate,ethanesulfonate, glucoheptanoate, glycerophosphate, hemisulfate,heptanoate, hexanoate, fumarate, hydrochloride, hydrobromide,hydroiodide, 2-hydroxyethanesulfonate, lactate, maleate,methanesulfonate, nicotinate, 2-napthalenesulfonate, oxalate, pamoate,pectinate, sulfate, 3-phenylpropionate, picrate, pivalate, propionate,succinate, tartrate, thiocyanate, p-toluenesulfonate and undecanoate.Also, the basic nitrogen-containing groups can be quaternized with suchagents as loweralkyl halides, such as methyl, ethyl, propyl, and butylchloride, bromides, and iodides; dialkyl sulfates like dimethyl,diethyl, dibutyl, and diamyl sulfates, long chain halides such as decyl,lauryl, myristyl and stearyl chlorides, bromides and iodides, aralkylhalides like benzyl and phenethyl bromides, and others. Water oroil-soluble or dispersible products are thereby obtained.

Examples of acids which may be employed to form pharmaceuticallyacceptable acid addition salts include such inorganic acids ashydrochloric acid, sulphuric acid and phosphoric acid and such organicacids as oxalic acid, maleic acid, succinic acid and citric acid. Basicaddition salts can be prepared in situ during the final isolation andpurification of the compounds of formula (I), or separately by reactingcarboxylic acid moieties with a suitable base such as the hydroxide,carbonate or bicarbonate of a pharmaceutically acceptable metal cationor with ammonia, or an organic primary, secondary or tertiary amine.Pharmaceutically acceptable salts include, but are not limited to,cations based on the alkali and alkaline earth metals, such as sodium,lithium, potassium, calcium, magnesium, aluminum salts and the like, aswell as nontoxic ammonium, quaternary ammonium, and amine cations,including, but not limited to ammonium, tetramethylammonium,tetraethylammonium, methylamine, dimethylamine, trimethylamine,triethylamine, ethylamine, and the like. Other representative organicamines useful for the formation of base addition salts includediethylamine, ethylenediamine, ethanolamine, diethanolamine, piperazineand the like.

Compounds of the present invention can be administered in a variety ofways including enteral, parenteral and topical routes of administration.For example, suitable modes of administration include oral,subcutaneous, transdermal, transmucosal, iontophoretic, intravenous,intramuscular, intraperitoneal, intranasal, subdural, rectal, and thelike.

In accordance with other embodiments of the present invention, there isprovided a composition comprising GSK3-inhibitor compound of the presentinvention, together with a pharmaceutically acceptable carrier orexcipient.

Suitable pharmaceutically acceptable excipients include processingagents and drug delivery modifiers and enhancers, such as, for example,calcium phosphate, magnesium stearate, talc, monosaccharides,disaccharides, starch, gelatin, cellulose, methyl cellulose, sodiumcarboxymethyl cellulose, dextrose, hydroxypropyl-β-cyclodextrin,polyvinylpyrrolidinone, low melting waxes, ion exchange resins, and thelike, as well as combinations of any two or more thereof. Other suitablepharmaceutically acceptable excipients are described in “Remington'sPharmaceutical Sciences,” Mack Pub. Co., New Jersey (1991), incorporatedherein by reference.

Pharmaceutical compositions containing GSK-3 inhibitor compounds of thepresent invention may be in any form suitable for the intended method ofadministration, including, for example, a solution, a suspension, or anemulsion. Liquid carriers are typically used in preparing solutions,suspensions, and emulsions. Liquid carriers contemplated for use in thepractice of the present invention include, for example, water, saline,pharmaceutically acceptable organic solvent(s), pharmaceuticallyacceptable oils or fats, and the like, as well as mixtures of two ormore thereof. The liquid carrier may contain other suitablepharmaceutically acceptable additives such as solubilizers, emulsifiers,nutrients, buffers, preservatives, suspending agents, thickening agents,viscosity regulators, stabilizers, and the like. Suitable organicsolvents include, for example, monohydric alcohols, such as ethanol, andpolyhydric alcohols, such as glycols. Suitable oils include, forexample, soybean oil, coconut oil, olive oil, safflower oil, cottonseedoil, and the like. For parenteral administration, the carrier can alsobe an oily ester such as ethyl oleate, isopropyl myristate, and thelike. Compositions of the present invention may also be in the form ofmicroparticles, microcapsules, liposomal encapsulates, and the like, aswell as combinations of any two or more thereof.

The compounds of the present invention may be administered orally,parenterally, sublingually, by inhalation spray, rectally, or topicallyin dosage unit formulations containing conventional nontoxicpharmaceutically acceptable carriers, adjuvants, and vehicles asdesired. Topical administration may also involve the use of transdermaladministration such as transdermal patches or ionophoresis devices. Theterm parenteral as used herein includes subcutaneous injections,intravenous, intramuscular, intrastemal injection, or infusiontechniques.

Injectable preparations, for example, sterile injectable aqueous oroleaginous suspensions may, be formulated according to the known artusing suitable dispersing or wetting agents and suspending agents. Thesterile injectable preparation may also be a sterile injectable solutionor suspension in a nontoxic parenterally acceptable diluent or solvent,for example, as a solution in 1,3-propanediol. Among the acceptablevehicles and solvents that may be employed are water, Ringer's solution,and isotonic sodium chloride solution. In addition, sterile, fixed oilsare conventionally employed as a solvent or suspending medium. For thispurpose any bland fixed oil may be employed including synthetic mono- ordiglycerides. In addition, fatty acids such as oleic acid find use inthe preparation of injectables.

Suppositories for rectal administration of the drug can be prepared bymixing the drug with a suitable nonirritating excipient such as cocoabutter and polyethylene glycols that are solid at ordinary temperaturesbut liquid at the rectal temperature and will therefore melt in therectum and release the drug.

Solid dosage forms for oral administration may include capsules,tablets, pills, powders, and granules. In such solid dosage forms, theactive compound may be admixed with at least one inert diluent such assucrose lactose or starch. Such dosage forms may also comprise, as isnormal practice, additional substances other than inert diluents, e.g.,lubricating agents such as magnesium stearate. In the case of capsules,tablets, and pills, the dosage forms may also comprise buffering agents.Tablets and pills can additionally be prepared with enteric coatings.

Liquid dosage forms for oral administration may include pharmaceuticallyacceptable emulsions, solutions, suspensions, syrups, and elixirscontaining inert diluents commonly used in the art, such as water. Suchcompositions may also comprise adjuvants, such as wetting agents,emulsifying and suspending agents, cyclodextrins, and sweetening,flavoring, and perfuming agents.

In accordance with yet other embodiments, the present invention providesmethods for inhibiting GSK3 activity in a human or animal subject, saidmethod comprising administering to a subject an amount of a GSK3inhibitor compound having the structure (I), (IV) or (V) (or compositioncomprising such compound) effective to inhibit GSK3 activity in thesubject. Other embodiments provided methods for treating a cell or aGSK3-mediated disorder in a human or animal subject, comprisingadministering to the cell or to the human or animal subject an amount ofa compound or composition of the invention effective to inhibit GSK3activity in the cell or subject. Preferably, the subject will be a humanor non-human animal subject. Inhibition of GSK3 activity includesdetectable suppression of GSK3 activity either as compared to a controlor as compared to expected GSK3 activity.

Effective amounts of the compounds of the invention generally includeany amount sufficient to detectably inhibit GSK3 activity by any of theassays described herein, by other GSK3 kinase activity assays known tothose having ordinary skill in the art or by detecting an alleviation ofsymptoms in a subject afflicted with a GSK3-mediated disorder.

GSK3-mediated disorders that may be treated in accordance with theinvention include any biological or medical disorder in which GSK3activity is implicated or in which the inhibition of GSK3 potentiatessignaling through a pathway that is characteristically defective in thedisease to be treated. The condition or disorder may either be caused orcharacterized by abnormal GSK3 activity. Representative GSK3-mediateddisorders include, for example, type 2 diabetes, Alzheimer's disease andother neurodegenerative disorders, obesity, atheroscleroticcardiovascular disease, essential hypertension, polycystic ovarysyndrome, syndrome X, ischemia, especially cerebral ischemia, traumaticbrain injury, bipolar disorder, immunodeficiency, cancer and the like.

Successful treatment of a subject in accordance with the invention mayresult in the inducement of a reduction or alleviation of symptoms in asubject afflicted with a medical or biological disorder to, for example,halt the further progression of the disorder, or the prevention of thedisorder. Thus, for example, treatment of diabetes can result in areduction in glucose or HbAlc levels in the patient. Likewise, treatmentof Alzheimer's disease can result in a reduction in rate of diseaseprogression, detected, for example, by measuring a reduction in the rateof increase of dementia.

The amount of active ingredient that may be combined with the carriermaterials to produce a single dosage form will vary depending upon thehost treated and the particular mode of administration. It will beunderstood, however, that the specific dose level for any particularpatient will depend upon a variety of factors including the activity ofthe specific compound employed, the age, body weight, general health,sex, diet, time of administration, route of administration, rate ofexcretion, drug combination, and the severity of the particular diseaseundergoing therapy. The therapeutically effective amount for a givensituation can be readily determined by routine experimentation and iswithin the skill and judgment of the ordinary clinician.

For purposes of the present invention, a therapeutically effective dosewill generally be from about 0.1 mg/kg/day to about 100 mg/kg/day,preferably from about 1 mg/kg/day to about 20 mg/kg/day, and mostpreferably from about 2 mg/kg/day to about 10 mg/kg/day of a GSK3inhibitor compound of the present invention, which may be administeredin one or multiple doses.

The compounds of the present invention can also be administered in theform of liposomes. As is known in the art, liposomes are generallyderived from phospholipids or other lipid substances. Liposomes areformed by mono- or multilamellar hydrated liquid crystals that aredispersed in an aqueous medium. Any non-toxic, physiologicallyacceptable and metabolizable lipid capable of forming liposomes can beused. The present compositions in liposome form can contain, in additionto a compound of the present invention, stabilizers, preservatives,excipients, and the like. The preferred lipids are the phospholipids andphosphatidyl cholines (lecithins), both natural and synthetic. Methodsto form liposomes are known in the art. See, for example, Prescott, Ed.,Methods in Cell Biology, Volume XIV, Academic Press, New York, N.W., p.33 et seq (1976).

While the compounds of the invention can be administered as the soleactive pharmaceutical agent, they can also be used in combination withone or more other agents used in the treatment of disorders.Representative agents useful in combination with the compounds of theinvention for the treatment of type 2 diabetes include, for example,insulin, troglitazone, rosiglitazone, pioglitazone, glipizide,metformin, sulfonylurea, acarbose, and the like. Representative agentsuseful in combination with the compounds of the invention for thetreatment of Alzheimer's disease include, for example, donepezil,tacrine and the like. Representative agents useful in combination withthe compounds of the invention for the treatment of bipolar diseaseinclude, for example, lithium salts, valproate, carbamazepine and thelike. A representative agent useful in combination with the compounds ofthe invention for the treatment of stroke is, for example, tissueplasminogen activator.

When additional active agents are used in combination with the compoundsof the present invention, the additional active agents may generally beemployed in therapeutic amounts as indicated in the PHYSICIANS' DESKREFERENCE (PDR) 53^(rd) Edition (1999), which is incorporated herein byreference, or such therapeutically useful amounts as would be known toone of ordinary skill in the art.

The compounds of the invention and the other therapeutically activeagents can be administered at the recommended maximum clinical dosage orat lower doses. Dosage levels of the active compounds in thecompositions of the invention may be varied so as to obtain a desiredtherapeutic response depending on the route of administration, severityof the disease and the response of the patient. The combination can beadministered as separate compositions or as a single dosage formcontaining both agents. When administered as a combination, thetherapeutic agents can be formulated as separate compositions that aregiven at the same time or different times, or the therapeutic agents canbe given as a single composition.

The foregoing and other aspects of the invention may be betterunderstood in connection with the following representative examples.

EXAMPLES Example 1 Characterization and Purification Methods

Compounds of the present invention were characterized by highperformance liquid chromatography (HPLC) using a Waters Millenniumchromatography system with a 2690 Separation Module (Milford, Mass.).The analytical columns were Alltima C-18 reversed phase, 4.6×250 mm fromAlltech (Deerfield, Ill.). A gradient elution was used, typicallystarting with 5% acetonitrile/95% water and progressing to 100%acetonitrile over a period of 40 minutes. All solvents contained 0.1%trifluoroacetic acid (TFA). Compounds were detected by ultraviolet light(UV) absorption at either 220 or 254 nm. HPLC solvents were from Burdickand Jackson (Muskegan, Mich.), or Fisher Scientific (Pittsburgh, Pa.).In some instances, purity was assessed by thin layer chromatography(TLC) using glass- or plastic-backed silica gel plates, such as, forexample, Baker-Flex Silica Gel 1B2-F flexible sheets. TLC results werereadily detected visually under ultraviolet light, or by employing wellknown iodine vapor and other various staining techniques.

Mass spectrometric analysis was performed on a Fisons VG ElectrosprayMass Spectrometer. All masses are reported as those of the protonatedparent ions.

Nuclear magnetic resonance (NMR) analysis was performed with a Varian300 MHz NMR (Palo Alto, Calif.). The spectral reference was either TMSor the known chemical shift of the solvent. Some compound samples wererun at elevated temperatures (i.e. 75° C.) to promote increased samplesolubility.

The purity of some of the invention compounds was assessed by elementalanalysis (Desert Analytics, Tucson, Ariz.)

Melting points were determined on a Laboratory Devices Mel-Tempapparatus (Holliston, Mass.).

Preparative separations were carried out using either a Flash 40chromatography system and KP-Sil, 60A (Biotage, Charlottesville, Va.), aChromatotron radial chromatography device (Harrison Research, Palo Alto,Calif.), or by HPLC using a C-18 reversed phase column. Typical solventsemployed were dichloromethane, methanol, ethyl acetate and triethylamine.

Example 2 Solid Phase Synthesis of Pyrimidine Compounds Resin Method A

Step A: Knoevenagel Condensation

A suspension of benzaldehyde-bound resin (1g, 0.52 mmol) in 8 ml of 1:1alcohol:dioxane was treated with 2.2 mole β-ketoester and 1.3 mmol anamine, e.g., piperidine. The reaction mixture was shaken for 20 hours atroom temperature and the resin was then filtered and washed with 4×10 mldichloromethane (DCM).

Step B: Cyclization and Oxidation to the Pyrimidine Nucleus

The product from Step A (100 mg, 0.052 mmol) was combined with 0.26 mmolof the pyrazole carboxamidine hydrochloride and 0.13 mmol NaHCO₃ in 1 mlN-methylpyrrolidinone. The reaction mixture was shaken at 70° C. for 24hours. Following cooling, the reaction was washed successively withwater, methanol, DMF, methylene chloride and ether, then dried. Cleavageof a small amount of resin indicated that the desired dihydropyrimidinewas present in high yield.

The dried resin was then taken up in THF and 1.1 eq. ofdicyanodichloroquinone (DDQ) was added. The resulting slurry was stirredfor 0.5 hours at which time the resin was washed with DMF, 10% Na₂HCO₃,H₂O, dimethylformamide (DMF), methanol (MeOH), methylene chloride andether, then dried. Cleavage of a small amount of this resin withtrifluoroacetic acid/methylene chloride indicated the presence of apyrimidine in high yield.

Step C: Amine Displacement and Release from the Solid Support

A suspension of the pyrimidine (50 mg, 0.026 mmol) in 0.75 ml NMP wastreated with 1 mmol of an amine and 0.26 mmol acetic acid. The reactionmixture was shaken at 80° C. for 24-48 hours. Following cooling, theresin was washed 4× each with methanol, DMF, and methylene chloride. Theresin was then dried and a solution of 5% trifluoroacetic acid inmethylene chloride was added. The resin was shaken for 2 hours, thenfiltered and washed 3× with methylene chloride. The combined filtrateswere concentrated, taken up in 1:1 water/acetonitrile and lyophilized todryness.

The following compounds of the present invention were prepared accordingto Resin Method A using the ketoester and amine identified inparentheses:

Ethyl4-(4-carbamoylphenyl)-6-ethyl-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate(from ethyl 3-oxo-valerate and 2-(2-aminoethylamino)-5-nitropyridine(dehydration of this compound using trifluoracetic anhydride yieldedethyl4-(4-cyanophenyl)-6-ethyl-2-({2-[(5-nitro(2-pyridyl)amino]ethyl}amino)pyrimidine-5-carboxylate)

Ethyl4-(4-carbamoylphenyl)-2-({2-[5-nitro(2-pyridyl))amino]ethyl}amino)-6-(4-pyridyl)pyrimidine-5-carboxylate(from ethyl 3-(4-pyridyl)-3-oxopropionate and2-(2-aminoethylamino)-5-nitropyridine)

Ethyl4-(4-carbamoylphenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-6-(4-nitrophenyl)pyrimidine-5-carboxylate(from ethyl 3-(4-nitrophenyl)-3-oxoproprionate and2-(2-aminoethylamino)-5-nitropyridine)

Example 3 Solid Phase Synthesis of Pyrimidine Compounds Resin Method B

Step 1: Sasrin resin (Bachem Biosciences, 5.0 g, nominal substitution1.02 mmol/g) was shaken with triphenylphosphine dibromide (2.3 g) in drydichloromethane (60-70 ml) for 4 hours at room temperature. All solventsand glassware used to carry out this reaction were dry. The resin waswashed well with dichloromethane.

Step 2: The resin from Step 1 was then reacted with a primary amine(0.5-1 M) in 1-methylpyrrolidone (NMP) at 70-80° C. for 3-5 hours toproduce an aminomethyl resin, which was used immediately afterpreparation. The resin was then thoroughly washed with dimethylsulfoxide(DMSO) (or DMF) and dichloromethane, then dried in vacuo at roomtemperature.

Step 3: After drying, the resin was coupled overnight with4-acetylbenzoic acid usingbenzotriazole-1-yl-oxy-tris-pyrollidino-phosphonium hexafluorophosphate(PyBop®, which is available from Novabiochem, San Diego, Calif.),4-methylmorpholine and NMP in accordance with the method described inExample 10 (i.e., “Resin Method C”) (except that cleavage of the productfrom the resin was carried out under more strongly acidic conditions,i.e., typically 20-100% trifluoroacetic acid (TFA) in DCM (e.g., 60% TFAin DCM)).

Other types of resin having a pendant CH₂OH group can also be used incarrying out this method such as, for example, Wang resin (Novabiochem,San Diego, Calif.). It is also possible to load the primary amine onto asolid support by other methods such as, for example, reductive aminationof a solid support containing an aldehyde.

Examples 4-9 describe the synthesis of compounds of the presentinvention pursuant to Resin Method B.

Example 4 Synthesis ofN-{(3-bromophenyl)methyl]{4-[2-({3-[(5-nitro(2-pyridyl)amino]propyl}amino)pyrimidin-4-yl]phenyl}carboxamide

Step 1: A solution of 2-chloro-5-nitropyridine (3.16 g, 20 mmol) in dryacetonitrile (40 ml) was added dropwise to a solution of1,3-diaminopropane (5.0 ml) in acetonitrile (20 ml) at room temperature.After 7.5 hour, a yellow solid precipitated in the reaction mixture. Thesolvent was removed in vacuo and the residue was partitioned between 2.5M aqueous sodium hydroxide and dichloromethane. The layers wereseparated and the aqueous portion was extracted 3× with dichloromethane.The combined organic layers were back-extracted with a saturated sodiumchloride solution, then dried and concentrated in vacuo using a Buchirotary evaporator Model R-124 to give (3-aminopropyl)(5-nitro(2-pyridyl))amine as a yellow solid (2.55 g). The amine (1.14 g, 6 mmol)was shaken with benzotriazole carboxamidinium 4-methylbenzenesulfonate(2.0 g, 6 mmol) and diisopropylethyl amine (DIEA) (1.05 ml, 6 mmol) inacetonitrile (10 ml) at room temperature over two days. Dilution withether gave the amino-{3-[(5-nitro(2-pyridyl))amino]propyl}carboxamidinium 4-methylbenzenesulfonate as asolid.

Step 2: Sasrin resin (10 g) was shaken with triphenylphosphine dibromide(4.5 g) in dry dichloromethane (ca. 80 ml) at room temperature for 4hours. The resin was washed well with dichloromethane and air-driedbriefly. The air-dried resin was divided into 6 equal portions. Oneportion was heated at 70° C. for 4 hours with a solution of3-bromobenzylamine (8 mmol) in NMP (12 ml). The resin was washed wellwith DMF and dichloromethane and dried overnight in vacuo at roomtemperature. The dried resin was then shaken with a solution of PyBop®(3.12 g, 6 mmol), 4-acetylbenzoic acid (1.0 g, 6 mmol), and4-methylmorpholine (12 mmol) in NMP (12 ml) at room temperatureovernight. The resin was washed with DMF, DMSO and dichloromethane andbriefly air-dried. The resin was then heated with N,N-dimethylformamidedimethylacetal (10 ml) at 95° C. for 9 hours. After cooling, the resinwas washed with dichloromethane and dried in vacuo at room temperature.The resin (80 mg) was then reacted with 100 mg of the guanidine preparedin Step 1 plus cesium carbonate (160 mg) with NMP (2 ml) at 95° C.overnight, followed by cleavage with 60% TFA in dichloromethane to giveN-{(3-bromophenyl)methyl]{4-[2-({3-[(5-nitro(2-pyridyl)amino]propyl}amino)pyrimidin-4-yl]phenyl}carboxamide.

HPLC: 25.31 min (98% pure); MS: MH⁺=562/564 (1 Br); C₂₆H₂₄N₇BrO₃=561/563g/mol.

Example 5 Synthesis ofN-{(3-bromophenyl)methyl]{4-[2-({2-[(5-cyano(2-pyridyl)aminolethyl}amino)pyrimidin-4-yl]phenyl}carboxamide

Step 1: 6-Chloronicotinitrile (2.0 g) was treated with ethylenediamine(5 ml). The mixture was then heated at 50° C. for 22 hours. Excessethylenediamine was removed by rotary evaporation. The residue waspartitioned between 2.5 M aqueous sodium hydroxide and dichloromethane.The aqueous layer was extracted 4× more with dichloromethane. Thecombined organic layers were washed with a saturated sodium chloridesolution, dried and then concentrated in vacuo to give6-[(2-aminoethyl)amino]pyridine-3-carbonitrile as an amber liquid whichsolidified upon standing. The amine (0.97 g, 6 mmol) was shakenovernight with benzotriazole carboxamidinium 4-methylbenzenesulfonate(2.0 g, 6 mmol) and DIEA (1.05 ml, 6 mmol) in acetonitrile (10 ml).Addition of ether gaveamino{2-[(5-cyano(2-pyridyl)amino]ethyl}carboxamidinium4-methylbenzenesulfonate as a white solid.

Step 2: The guanidine from Step 1 (120 mg) was reacted with the resinprepared as in Example 4, Step 2 (80 mg) in the presence of cesiumcarbonate (160 mg) in NMP (2 ml) at 90° C. overnight. Treatment of theresin with 60% TFA in dichloromethane gave the title compound.

HPLC: 23.70 min (98% purity)

MS: MH⁺=528/530 (1 Br) C₂₆H₂₂N₇BrO=527/529 g/mole

Example 6 Synthesis ofN-[(3-methoxyphenyl)methyl]{4-[2-({2-[(5-nitro(2-pyridyl))aminolethyl}amino)pyrimidin-4-yl]phenyl}carboxamide

Step 1: 2-(2-aminoethylamino)-5-nitropyridine (Aldrich Chemical Co.,Milwaukee, Wis.) (1.08 g, 6 mol) was shaken with benzotriazolecarboxamidinium 4-methylbenzenesulfonate (2.0 gm, 6 mmol) and DIEA (1.05ml, 6 mmol) in a mixture of acetonitrile (10 ml) and DMF (3 ml) at roomtemperature overnight. Addition of ether gaveamino[2-[(5-nitro(2-pyridyl)amino]ethyl}carboxamidinium4-methylbenzene-sulfonate as light orange crystals.

Step 2: The Sasrin resin was prepared as described in Step 2 of Example3. The resin (500 mg) was heated with a solution of 3-methoxybenzylamine(600 μl) in NMP (6 ml) at 70° C. for 4 hours. The resin was then washedwith DMF and dichloromethane and then shaken with a solution of PyBop®(1.04 g, 2 mmol), 4-acetylbenzoic acid (0.33 g, 2 mmol),4-methylmorpholine (4 mmol) in NMP (6 ml) at room temperature overnight.A small aliquot of the resin was treated with 20% TFA in dichloromethaneto give the intermediate,(4-acetylphenyl)-N-[(3-methoxyphenyl)methyl]carboxamide (HPLC: 23.94 min(97%); MS; MH⁺=284 (as required)). The resin was then heated at 95° C.for 7 hours in DMFDMA (5 ml).

After heating, the resin was washed with dichloromethane, then dried invacuo. Dried resin (120 mg) was reacted with 120 mg of the guanidineprepared in Step 1 plus cesium carbonate (160 mg) in NMP (2 ml)overnight at 90° C. Cleavage with 20% TFA in dichloromethane gave thetitle compound.

HPLC: 22.32 min (85% pure)

MS: MH⁺=500 C₂₆H₂₅N₇O₄=499 g/mole

Example 7 Synthesis of4-(2-{[3-(4-nitroimidazolyl)propyl]amino}pyrimidin-4-yl)phenol

Step 1: 4-Nitroimidazole (5.0 g, 44 mol) in DMF:THF (1:1 (v/v), 40 ml)was treated at room temperature with 60% NaH (2.2 g). When hydrogenevolution had ceased, 3-bromopropylphthalimide (11.79 g, 44 mmol) wasadded, followed by heating at 70° C. overnight. The mixture was cooled,diluted with dichloromethane and carefully quenched with water. At thispoint the solid product precipitated out to give2-[3-(4-nitroimidazolyl)propyl]isoindoline-1,3-dione as a white solid,8.85 g. The solid was refluxed with methanol (60 ml) and anhydroushydrazine (4 ml) overnight. The mixture was cooled to 4° C., thenfiltered. The filtrate was concentrated to dryness, then partitionedbetween dichloromethane and 2.5 M aqueous sodium hydroxide. The organiclayer was washed with saturated sodium chloride solution, dried andconcentrated in vacuo to give 3-(4-nitroimidazolyl)propylamine as anorange syrup, 2.24 g. The amine, 1.18 g was treated with benzotriazolecarboxamidinium 4-methylbenzenesulfonate (2.2 g) and DIEA (1.5 ml) inacetonitrile (8 ml) with shaking at room temperature overnight. Additionof ether gave amino[3-(4-nitroimidazolyl)propyl]carboxamidinium4-methylbenzenesulfonate as a beige solid.

Step 2: The Sasrin resin (prepared according to Example 3, Step 2) (2.5g) was heated at 80° C. with 4-hydroxyacetophenone (700 mg) and cesiumcarbonate (600 mg) in NMP (10 ml) for 24 hours. The resin was thenwashed with DMF, water, DMF and dichloromethane and dried in vacuo. Thedried resin was then heated overnight with DMFDMA (10 ml) at 105° C. Theresin was cooled, filtered and washed well with dichioromethane anddried in vacuo. The dried resin (100 mg) was then treated with 100 mg ofthe guanidine prepared in Step 1, 200 mg of cesium carbonate and 3 ml ofNMP at 105° C. for 66 h. The resin was washed with DMSO, acetic acid,water, DMSO and dichloromethane, then shaken with 100% TFA for 1 h, andfiltered. The filtrate was concentrated in vacuo and lyophilized to givethe title compound.

HPLC: 16.85 min (75% purity)

MS: MH⁺=341 C₁₆H₁₆N₆O₃=340 g/mol

Example 8 Synthesis of4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-5-phenylpyrimidin-4-yl]phenol

Bromomethyl sasrin resin (prepared according to Example 3, Step 2), 0.9g, was heated with benzyl 4-hydroxyphenyl ketone (1.06 g, 5 mmol) andcesium carbonate (1.6 g) in NMP (8 ml) at 80° C. overnight. The resinwas washed serially with DMF, water, DMF and dichloromethane and driedin vacuo. The dried resin was heated with DMFDMA (8 ml) at 100° C.overnight. After cooling the resin was filtered and washed well withdichloromethane, then dried in vacuo. The resin (75 mg) was then reactedwith 100 mg of amino {2-[(5-nitro(2-pyridyl))amino]ethyl}carboxamidinium4-methylbenzenesulfonate and 200 mg of cesium carbonate in NMP (2 ml) at104° C. for 64 hours. The resin was then washed with DMSO, acetic acid,water, DMSO and dichloromethane. The resin was shaken with 100% TFA atroom temperature (1 h). The resin was filtered and the filtrateconcentrated in vacuo, then lyophilized to give the title compound.

HPLC: 22.53 min (95% purity)

MS: MH⁺=429 C₂₃H₂₀N₆O₃=428 g/mol

Example 9 Synthesis of[(3-Bromophenyl)methyl]({4-[2-({2-[(5-nitro(2-pyridyl))aminolethyl}amino)pyrimidine-4-yl]phenyl}sulfonyl)amine

Step 1: Sasrin resin (500 mg) substituted with m-bromobenzylamine(according to Step 1 of Example 3) was treated with4-acetylbenzenesulfonyl chloride (1.1 g, 5 mmol) and DIEA (1.22 ml, 7mmol) in dichloromethane (10 ml) with shaking at room temperature for0.5 hours. Then 4-dimethylaminopyridine (122 mg, 1 mmol) was added,followed by shaking overnight at room temperature. The resin was washedwell with DMF and dichloromethane, then heated with DMFDMA (10 mL) at95° C. overnight. The resin was washed well with dichloromethane anddried in vacuo at room temperature.

Step 2: The resin prepared in Step 1 (70 mg) was treated withamino[2-[(5-nitro(2-pyridyl)amino]ethyl}carboxamidinium4-methylbenzenesulfonate (100 mg) and cesium carbonate (160 mg) in NMP 2ml) at 95° C. overnight. The resin was serially washed with DMSO, aceticacid, water, DMSO, dichloromethane and then treated with 60% TFA indichloromethane at room temperature for 0.5 hours. The resin wasfiltered off and the filtrate was concentrated in vacuo and lyophilizedto give the title compound.

HPLC: 26.62 min (100% purity)

MS: MH⁺=584/586 C₂₄H₂₂N₇BrO₄S=583/585 g/mol (1 Br)

The following additional compounds were similarly synthesized accordingto Resin Method B by varying the guanidine used:

-   4-(2-{[2-(4-nitrophenyl)ethyl]amino}pyrimidin-4-yl)benzamide-   4-{2-[(2-(2-pyridyl)ethyl)amino]pyrimidin-4-yl}benzamide-   4-{2-[(4-pyridylmethyl)amino]pyrimidin-4-yl}benzamide-   4-[2-({2-[(5-nitro-2-pyridyl)amino]ethyl}amino)pyrimidin-4-yl]benzamide-   4-(2-{[2-(pyrimidin-2-ylamino)ethyl]amino}pyrimidin-4-yl)benzamide-   4-{2-[(3-imidazol-5-ylethyl)amino]pyrimidin-4-yl}benzamide-   4-(2-{[2-(benzothiazol-2-ylamino)ethyl]amino}pyrimidin-4-yl)benzamide-   4-{2-[(2-{[5-(trifluoromethyl)-2-pyridyl]amino}ethyl)amino]pyrimidin-4-yl}benzamide-   4-[2-({2-[(5-cyano-2-pyridyl)amino]ethyl}amino)pyrimidin-4-yl]benzamide-   4-{2-[(2-{[5-(aminothioxomethyl)-2-pyridyl]amino}ethyl)amino]pyrimidin-4-yl}benzamide-   4-(2-{[(3-bromophenyl)methyl]amino}pyrimidin-4-yl)benzamide-   4-[2-({[4-(4-fluorophenyl)phenyl]methyl}amino)pyrimidin-4-yl]benzamide-   4-{2-[4-benzylpiperazinyl]pyrimidin-4-yl}benzamide-   4-(2-{[(5-methylpyrazin-2-yl)methyl]amino}pyrimidin-4-yl)benzamide-   4-{2-[(3-imidazolylpropyl)amino]pyrimidin-4-yl}benzamide-   4-{2-[(2,2-diphenylethyl)amino]pyrimidin-4-yl}benzamide-   4-[2-({[3-(trifluoromethyl)phenyl]methyl}amino)pyrimidin-4-yl]benzamide-   4-(2-{[(3-nitrophenyl)methyl]amino}pyrimidin-4-yl)benzamide-   4-{2-[(naphthylmethyl)amino]pyrimidin-4-yl}benzamide-   4-(2-{[(4-bromophenyl)methyl]amino}pyrimidin-4-yl)benzamide-   4-(2-{[(3,5-dichlorophenyl)methyl]amino}pyrimidin-4-yl)benzamide-   4-(2-{[(3-methoxyphenyl)methyl]amino}pyrimidin-4-yl)benzamide-   4-[2-({[3-(3-methoxyphenyl)phenyl]methyl}amino)pyrimidin-4-yl]benzamide-   4-[2-({[3-(3-aminophenyl)phenyl]methyl}amino)pyrimidin-4-yl]benzamide-   4-{2-[({3-[3-(acetylamino)phenyl]phenyl{methyl)amino]pyrimidin-4-yl}benzamide-   4-[2-({[4-(3-aminophenyl)phenyl]methyl}amino)pyrimidin-4-yl]benzamide-   4-(2-{[(3-chlorophenyl)methyl]amino}pyrimidin-4-yl)benzamide-   4-(2-{[(2,4-dichlorophenyl)methyl]aminol}pyrimidin-4-yl)benzamide-   4-(2-{[(3-methylphenyl)methyl]amino}pyrimidin-4-yl)benzamide-   4-(2-{[(3,4-dimethoxyphenyl)methyl]amino}pyrimidin-4-yl)benzamide-   4-[2-({[4-(trifluoromethyl)phenyl]methyl}amino)pyrimidin-4-yl]benzamide-   4-(2-{[(4-methoxyphenyl)methyl]amino}pyrimidin-4-yl)benzamide-   4-(2-{[(4-aminophenyl)methyl]amino}pyrimidin-4-yl)benzamide-   4-[2-({[3,5-bis(trifluoromethyl)phenyl]methyl}amino)pyrimidin-4-yl]benzamide-   4-{2-[4-(2-methoxyphenyl)piperazinyl]pyrimidin-4-yl}benzamide-   4-{2-[({3-[3-(trifluoromethyl)phenyl]phenyl}methyl)amino]pyrimidin-4-yl}benzamide-   5 4-(2-{[2-(3-methoxyphenyl)ethyl]amino}pyrimidin-4-yl)benzamide-   4-(2-{[2-(4-fluorophenyl)ethyl]amino}pyrimidin-4-yl)benzamide-   4-(2-{[(3,4,5-trimethoxyphenyl)methyl]amino}pyrimidin-4-yl)benzamide-   {4-[2-({2-[(4-amino-5-cyanopyrimidin-2-yl)amino]ethyl}amino)pyrimidin-4-yl]phenyl}-N-[(3-bromophenyl)methyl]carboxamide-   4-[2-({2-[(4-amino-5-cyanopyrimidin-2-yl)amino]ethyl}amino)pyrimidin-4-yl]benzamide-   4-[2-({3-[(5-nitro-2-pyridyl)amino]propyl}amino)pyrimidin-4-yl]benzamide-   4-(2-{[(4-cyanophenyl)methyl]amino}pyrimidin-4-yl)benzamide-   4-2-[(2-phenylpropyl)amino]pyrimidin-4-yl}benzamide-   4-{2-[(2-phenoxyethyl)amino]pyrimidin-4-yl}benzamide-   4-(2-{[2-(2,5-dimethoxyphenyl)ethyl]amino}pyrimidin-4-yl)benzamide-   4-(2-{[(2,6-dimethoxyphenyl)methyl]amino}pyrimidin-4-yl)benzamide-   4-(2-{[2-(2-methoxyphenyl)ethyl]amino}pyrimidin-4-yl)benzamide-   4-{2-[(4-phenylbutyl)amino]pyrimidin-4-yl}benzamide-   4-{2-[(2-(2H-benzo[3,4-d]1,3-dioxolen-5-yl)ethyl)amino]pyrimidin-4-yl}benzamide-   4-(2-{[2-(3-chlorophenyl)ethyl]amino}pyrimidin-4-yl)benzamide-   4-(2-{[2-(2,3-dimethoxyphenyl)ethyl]amino}pyrimidin-4-yl)benzamide-   4-{2-[(3-phenoxypropyl)amino]pyrimidin-4-yl}benzamide-   4-(2-{[3-(4-chlorophenoxy)propyl]amino}pyrimidin-4-yl)benzamide-   4-(2-{[3-(4-methoxyphenoxy)propyl]amino}pyrimidin-4-yl)benzamide-   4-(2-{[3-(3-methoxyphenoxy)propyl]amino}pyrimidin-4-yl)benzamide-   4-(2-{[3-(3-bromophenoxy)propyl]amino}pyrimidin-4-yl)benzamide-   4-(2-{[3-(2,4-dichlorophenoxy)propyl]amino}pyrimidin-4-yl)benzamide-   4-[2-({3-[3-(trifluoromethyl)phenoxy]propyl}amino)pyrimidin-4-yl]benzamide-   4-(2-{[3-(3-methylphenoxy)propyl]amino}pyrimidin-4-yl)benzamide-   4-(2-{[3-(4-phenylimidazolyl)propyl]amino}pyrimidin-4-yl)benzamide-   4-(2-{[3-(5,6-dichlorobenzimidazolyl)propyl]amino}pyrimidin-4-yl)benzamide-   4-(2-{[3-(3,4-dichlorophenoxy)propyl]amino}pyrimidin-4-yl)benzamide-   4-(2-{[3-(5,6-dimethylbenzimidazolyl)propyl]amino}pyrimidin-4-yl)benzamide-   4-{2-[(3-(6-quinolyloxy)propyl)amino]pyrimidin-4-yl}benzamide-   4-{2-[(3-naphthyloxypropyl)amino]pyrimidin-4-yl}benzamide-   4-(2-{[3-(3-phenylphenoxy)propyl]amino}pyrimidin-4-yl)benzamide-   4-(2-{[3-(4-nitroimidazolyl)propyl]amino}pyrimidin-4-yl)benzamide-   4-(2-{[3-(4,5-dichloroimidazolyl)propyl]amino}pyrimidin-4-yl)benzamide-   4-{2-[(3-benzimidazolylpropyl)amino]pyrimidin-4-yl}-2-chlorophenol-   4-[2-({3-[4-(2,4-dichlorophenyl)imidazolyl]propyl}amino)pyrimidin-4-yl]benzamide-   4-[2-({3-[4-(3-methoxyphenyl)imidazolyl]propyl}amino)pyrimidin-4-yl]benzamide

Example 10 Solid Phase Synthesis of Pyridine Compounds Resin Method C

Rink amide resin (Novabiochem, San Diego, Calif., nominally 0.46 mmol/gsubstitution) was deprotected with 20% v/v piperidine in DMF (ca. 60 ml,0.5 hours, room temperature). The resin was washed thoroughly with DMFand dichloromethane, then treated with 4-acetylbenzoic acid (8 mmol),PyBOP® (8 mmol, Novabiochem), 4-methylmorpholine (12 mmol) and NMP (50ml) for 8.5hours at room temperature on a wrist shaker. The resin waswashed with DMF and dichloromethane, air dried, and then divided into 3portions. Each portion was treated with N,N-dimethylformamide dimethylacetal (ca. 12 ml) with heating at 105° C. overnight (ca. 13 hours). Thereactions were allowed to cool and the resin was washed withdichloromethane, then dried in vacuo at room temperature.

For the synthesis of pyrimidines, typically 100 mg of the above driedresin was mixed with 200-300 mg of anhydrous cesium carbonate, 80-200 mg(most usually 100 mg) of the appropriate guanidine as its tosylate saltand 2-3 ml of NMP. This mixture was heated at 90-105° C. for at least 12hours. In many cases the reactions were allowed to proceed for about 65hours at this temperature. The resin was cooled, filtered and washedwith DMSO, glacial acetic acid, water, DMSO and finally dichloromethane.The product was removed by treatment of the resin with 95:5 v/vdichloromethane/TFA for 0.5-1 hours at room temperature. The resin wasthen filtered, washed with dichloromethane and the filtrates wereconcentrated on a rotary evaporator. An aliquot was withdrawn for HPLCanalysis and the rest of the sample was lyophilized twice from a 1:1acetonitrile:water solvent mixture, which usually gave the pyrimidine asa fluffy solid.

Examples 11-16 describe the synthesis of compounds of the presentinvention pursuant to Resin Method C.

Example 11 Synthesis of4-(2-{[2-(2-pyridylamino)ethyl]amino}pyrimidin-4-yl)benzamide

2-(2-Aminoethylamino)pyridine (prepared from 2-chloropyridine andethylenediamine in accordance with the method described in T. Mega etal., 1988, Bull. Chem. Soc. Japan 61:4315, which is incorporated hereinby reference) (6 mmol) was treated with benzotriazole carboxamidinium4-methylbenzenesulfonatesulfonate (2.0 g, 6 mmol) and DIEA (1.05 ml, 6mmol) in anhydrous acetonitrile (10 ml) for 65 hours. Ether (ca. 10 ml)was then added to this mixture. After 8 hours the white solidamino[2-(2-pyridylamino)ethyl]carboxamidinium 4-methylbenzenesulfonatewas filtered off and dried in vacuo. The resulting guanidine (200 mg)was reacted with 100 mg of resin (21 hours, 90° C.) according to themethod described in Example 10 (Resin method C), to give the titlecompound.

HPLC: 11.20 min (97% pure)

MS: MH⁺=335 C₁₈H₁₈N₆O=334 g/mol

Example 12 Synthesis of4-(2-{[2-(2-guinolylamino)ethyl]amino}pyrimidin-4-yl)benzamide

2-Chloroquinoline (7.0 g) was heated at 120° C. under argon withethylenediamine (50 ml) for 6 hours. The excess ethylenediamine wasremoved by rotary evaporation (oil pump). The residue was taken up in2.5 M aqueous sodium hydroxide and extracted 6 times withdichloromethane. The combined organic layers were washed with a smallportion of saturated aqueous NaCl, dried over Na₂SO₄ and concentrated invacuo. A portion of the viscous product, 0.55 g (3 mmol) was treatedwith benzotriazole carboxamidinium 4-methylbenzenesulfonate (1.0 g, 3mmol), DIEA (0.78 ml, 4.5 mmol) and acetonitrile (8 ml) with shaking atroom temperature overnight. Precipitation with ether gaveamino[2-(2-quinolylamino)ethyl]carboxamidinium 4-methylbenzenesulfonate.The resulting guanidine (200 mg) was reacted (21 hours, 90° C.) with 100mg of resin according to the method described in Example 10 (i.e., Resinmethod C) to give the title compound.

HPLC: 12.04 min (95% pure)

MS: MH⁺=385 C₂₂H₂₀N₆O=384 g/mol

Example 13 Synthesis of4-[2-({2-[6-methoxy-2-pyridyl)amino[-ethyl}amino)pyrimidin-4-yl]benzamide

2-Chloro-6-methoxypyridine (5.0 g) was heated with ethylenediamine (30ml) at 120° C. overnight. The excess ethylenediamine was removed byrotary evaporation. The residue was dissolved in a small volume of 2.5 Maqueous sodium hydroxide and extracted thoroughly with dichloromethane.The combined organic layers were washed with saturated aqueous sodiumchloride, dried over sodium sulfate and concentrated in vacuo to give(2-aminoethyl)(6-methoxy(2-pyridyl)amine as an orange syrup. The amine(2.58 g) was treated with benzotriazole carboxamidinium4-methylbenzenesulfonate (0.86 g) and DIEA (0.45 mmol) in acetonitrile(6 ml) and stirred overnight at room temperature. Trituration with ethergave the guanidine,amino-{2-[(6-methoxy(2-pyridyl))amino]ethyl}carboxamidinium4-methylbenzenesulfonate, as an oil. The oily guanidine (200 mg) wasreacted with 100 mg of resin according to Resin Method C (90° C.,overnight) to give the title compound.

HPLC: 11.84 min (85% purity)

MS: MH⁺=365 C₁₉H₂₀N₆O₂=364 g/mol

Example 14 Synthesis of4-{2-[(3-Benzimidazolylpropyl)amino]pyrimidin-4-yl]benzamide

Benzimidazole (2.4 g, 20 mmol) in dry THF (40 ml) was treated at roomtemperature with 60% NaH in oil (0.96 g). After hydrogen evolutionceased, 3-bromopropylphthalimide (5.36 g, 20 mmol) was added and themixture heated at 80° C. overnight. The reaction was cooled, dilutedwith dichloromethane and water and then extracted twice with 5% aqueouspotassium carbonate solution. The organic layer was dried over sodiumsulfate and concentrated in vacuo to give a beige solid, 4.1 g. Thesolid was dissolved in methanol (60 ml) and treated with anhydroushydrazine (4.0 ml), followed by refluxing for 4 hours. The mixture wasthen cooled to 4° C. for several hours, then filtered. The filtrate wasconcentrated in vacuo. The residue was partitioned betweendichloromethane and 2.5 M aqueous sodium hydroxide. The organic layerwas washed with saturated sodium chloride solution, dried over sodiumsulfate and concentrated in vacuo to give 3-benzimidazolylpropyl amineas a pale pink oil, 1.1 g. This amine (1.03 g, 6 mmol) was reacted withbenzotriazole carboxamidinium 4-methylbenzenesulfonate (2.0 g, 6 mmol)and DIEA (1.39 ml) in acetonitrile (8 ml) overnight at room temperatureto give amino(3-benzimidazolylpropyl)carboxamidinium4-methylbenzenesulfonate which was obtained as a beige solid afterrepeated trituration with ether. The guanidine (100 mg) was reacted) (65hours, 105° C.) with 100 mg of resin in accordance with the methoddescribed in Example 10 (i.e., Resin Method C) to give the titlecompound.

HPLC: 12.12 min (95% purity)

MS: MH⁺=373 C₂₁H₂₀N₆O=372 g/mol

Example 15 Synthesis of4-{2-[(3-(2-napthyloxy)propyl)amino]pyrimidin-4-yl)benzamide

2-Naphthol (2.9 g, 20 mmol) in dry THF (40 ml) was treated with 60% NaHsuspension (0.96 g) at room temperature. After hydrogen evolutionceased, 3-bromopropylphthalimide (5.36 g, 20 mmol) was added and themixture was heated at 80° C. overnight. The reaction was cooled, dilutedwith ethyl acetate and water. The layers were separated and the aqueouslayer extracted 3 times with ethyl acetate. The combined organic layerswere then extracted 5 times with 5% aqueous potassium carbonate, driedover sodium sulfate and concentrated in vacuo. The crude product(observed as a single spot by TLC) was taken up in methanol (60 ml),treated with anhydrous hydrazine (4 ml) and refluxed for 3.5 hours. Themixture was cooled to 4° C. for several hours, then filtered. Thefiltrate was concentrated to dryness, then partitioned betweendichloromethane and 2.5 M aqueous sodium hydroxide. The organic layerwas washed with saturated sodium chloride solution, dried andconcentrated in vacuo to give 3-(2-naphthyloxy)propyl amine as a beigesolid, 1.14 g. The amine (1.14 g, 5.7 mmol) was treated withbenzotriazole carboxamidinium 4-methylbenzenesulfonate (1.89 g, 5.7mmol) and DIEA (1.39 ml) in a mixture of acetonitrile (8 ml) and DMF (2ml) with shaking at room temperature overnight. Precipitation with ethergave amino(3-(2-naphthyloxy)propyl)carboxamidinium4-methylbenzenesulfonate as a white crystalline solid. This guanidine(100 mg) was reacted (65 hours, 105° C.) with resin (100 mg) inaccordance with the method described in Example 10 (i.e., Resin MethodC) to give the title compound.

HPLC: 22.52 min (95% purity)

MS: MH⁺=399 C₂₄H₂₂N₄O₂=398 g/mol

Example 16 Synthesis ofN-(1-carbamoyl-2-phenylethyl)(4-{2-{(2-(2-pyridyl)ethyl)amino]pyrimidin-4-yl}phenyl)carboxamide

This Example provides a variation of Resin Method C in which thepyrimidine is linked to an α-amino acid residue.

Step 1: Rink amide resin (1.5 g) was deprotected with 20% piperidine inDMF (1×0.5 hours). The resin was washed well with DMF and then treatedwith FMOC (L)-phenylalanine (5.0 mmol), 1-hydroxybenzotriazole (5.0mmol) and diisopropylcarbodiimide (5.0 mmol) in DMF (10 ml) with shakingat room temperature for 2 hours. The resin was washed with DMF and thentreated with 20% piperidine in DMF (1×30 min). The resin was washed wellwith DMF and then treated with PyBOP® (5 mmol), 4-methylmorpholine (8mmol) and 4-acetylbenzoic acid (5 mmol) in NMP (10 ml). After 5 hours atroom temperature, a negative ninhydrin test indicated completion of thereaction. The resin was washed with DMF and dichloromethane, air-driedand then heated with DMFDMA at 110° C. overnight. The resin was thenwashed well with dichloromethane and dried in vacuo at room temperature.

Step 2: The resin prepared in Step 1 (150 mg) was treated withamino(2-(2-pyridyl)ethyl)carboxamidinium 4-methylbenzenesulfonate (200mg) and cesium carbonate (160 mg) in NMP (2 ml) at 85° C. overnight. Theresin was washed with DMF and dichloromethane and then treated with 5%TFA in dichloromethane. The resin was filtered off and the filtrateconcentrated and lyophilized to give the title compound.

HPLC: 15.08 min (95% purity)

MS: MH⁺=467 C₂₇H₂₆N₆O₂=466 g/mol

The following additional compounds were analogously synthesizedaccording to Resin Method C by varying the guanidine used:

-   N-benzyl(4-{2-[(2-(2-pyridyl)ethyl)amino]pyrimidin-4-yl}phenyl)carboxamide-   benzyl    {[4-(2-{[2-(2-pyridylamino)ethyl]amino}pyrimidin-4-yl)phenyl]sulfonyl}amine-   {4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}-N-benzylcarboxamide-   {4-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}-N-benzylcarboxamide-   N-[(4-fluorophenyl)methyl]{4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}carboxamide-   N-[(3-bromophenyl)methyl]{4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}carboxamide-   N-(2-methoxyethyl)    {4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}carboxamide-   N-(naphthylmethyl){4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}carboxamide-   {4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}-N-{[3-(trifluoromethyl)phenyl]methyl}carboxamide-   {4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}-N-(2-phenylethyl)carboxamide-   N-[(4-methoxyphenyl)methyl]{4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}carboxamide-   N-[(3-methoxyphenyl)methyl]{4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}carboxamide-   {4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}-N-(oxolan-2-ylmethyl)carboxamide-   N-[(5-methylpyrazin-2-yl)methyl]{4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}carboxamide-   N-(2,2-diphenylethyl){4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}carboxamide-   {4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}-N-(4-piperidylmethyl)carboxamide-   N-[2-(2,4-dichlorophenyl)ethyl]{4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}carboxamide-   {4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}-N-(3-pyridylmethyl)carboxamide-   N-(3-imidazolylpropyl){4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}carboxamide-   {4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}-N-(2-thienylmethyl)carboxamide-   {4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}-N-[(3-nitrophenyl)methyl]carboxamide-   N-[(3-methylphenyl)methyl]{4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}carboxamide-   {4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}-N-[(4-sulfamoylphenyl)methyl]carboxamide-   {4-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}-N-[(3-bromophenyl)methyl]carboxamide-   N-[(3,5-dichlorophenyl)methyl]{4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}carboxamide-   N-[(3,4-difluorophenyl)methyl]{4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}carboxamide-   N-[(4-bromophenyl)methyl]{4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}carboxamide-   N-[(2,3-dimethoxyphenyl)methyl]{4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}carboxamide-   N-[(3-fluorophenyl)methyl]{4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}carboxamide-   N-[(3-bromophenyl)methyl]{4-[2-({2-[(6-methoxy(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}carboxamide-   [4-(2-{[(3-bromophenyl)methyl]amino}pyrimidin-4-yl)phenyl]-N-[(3-methylphenyl)methyl]carboxamide-   N-[(3-bromophenyl)methyl]{4-[2-({2-[(5-cyano(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}carboxamide-   4-(2-{[(3-{3-[(methylamino)methyl]phenyl}phenyl)methyl]amino}pyrimidin-4-yl)benzamide-   N-[(3-bromophenyl)methyl](4-{2-[(3-imidazolylpropyl)amino]pyrimidin-4-yl}phenyl)carboxamide-   N-[(3-bromophenyl)methyl][4-(2-{[2-(2-quinolylamino)ethyl]amino}pyrimidin-4-yl)phenyl]carboxamide-   N-[(3-bromophenyl)methyl]{4-[2-({2-[(4-nitrophenyl)amino]ethyl}amino)pyrimidin-4-yl]phenyl}carboxamide-   N-[(3-bromophenyl)methyl](4-{2-[(2-{[5-(trifluoromethyl)(2-pyridyl)]amino}ethyl)amino]pyrimidin-4-yl}phenyl)carboxamide-   N-[(3-bromophenyl)methyl][4-(2-{[2-(pyrimidin-2-ylamino)ethyl]amino}pyrimidin-4-yl)phenyl]carboxamide-   N-[(3-chlorophenyl)methyl]{4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}carboxamide-   N-[(3,4-dimethoxyphenyl)methyl]{4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}carboxamide-   N-[(3,4-dichlorophenyl)methyl]{4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}carboxamide-   [(3-bromophenyl)methyl]({4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}sulfonyl)amine-   N-[(3-iodophenyl)methyl]{4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}carboxamide-   [4-(2-{[2-(2,5-dimethoxyphenyl)ethyl]amino}pyrimidin-4-yl)phenyl]-N-[(3-bromophenyl)methyl]carboxamide-   N-[(3-bromophenyl)methyl][4-(2-{[2-(3-methoxyphenyl)ethyl]amino}pyrimidin-4-yl)phenyl]carboxamide-   {4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}-N-(2-phenylcyclopropyl)carboxamide-   3-[2-({2-[(5-nitro-2-pyridyl)amino]ethyl}amino)pyrimidin-4-yl]phenol-   4-[2-({2-[(5-nitro-2-pyridyl)amino]ethyl}amino)pyrimidin-4-yl]phenol-   4-{2-[(3-phenoxypropyl)amino]pyrimidin-4-yl}phenol-   4-(2-{[3-(4-chlorophenoxy)propyl]amino}pyrimidin-4-yl)phenol-   4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-5-phenylpyrimidin-4-yl]phenol-   4-{2-[(3-benzimidazolylpropyl)amino]pyrimidin-4-yl}phenol-   4-{2-[(3-benzimidazolylpropyl)amino]pyrimidin-4-yl}-2-methoxyphenol-   4-(2-{[3-(4-nitroimidazolyl)propyl]amino}pyrimidin-4-yl)phenol-   4-(2-[3-(2-aminobenzimidazolyl)propyl]amino}pyrimidin-4-yl)phenol-   4-(2-{[3-(4,5-dichloroimidazolyl)propyl]amino}pyrimidin-4-yl)phenol

Example 17 Solid Phase Synthesis of Pyrimidine Compounds Resin Method D

A primary amine was loaded onto Sasrin resin as in Example 3 (i.e.,Resin Method B). This amine resin was then heated with either2,4-dichloropyrimidine or ethyl 2,4-dichloropyrimidine-5-carboxylate(200 mg of pyrimidine per 200 mg of amine resin) and cesium carbonate(250 mg) in NMP (3 ml) overnight. The resin was washed with theappropriate solvents (typically DMF or DMSO and dichloromethane) andthen reacted with a second amine (e.g., a primary or secondary amine).Second amine displacement was typically conducted at a highertemperature in NMP, for example for 48 hours at 120-130° C. The resinwas again washed and treated with 100% TFA for 0.5-1 hours to obtain the2,4-diaminopyrimidine, which was frequently obtained as a solid afterlyophilization from a mixture of acetonitrile and water.

Examples 18-19 describe the synthesis of compounds of the presentinvention pursuant to Resin Method D.

Example 18 Synthesis of[(3-chlorophenyl)methyl)[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]amine

Bromomethyl Sasrin resin (prepared as in Step 1 of Example 3, 0.9 g) washeated with 3-chlorobenzylamine (1 ml) in NMP (8 ml) at 80° C. for 1.5hours, then overnight at room temperature. The resin was washed with DMFand dichloromethane and dried in vacuo. The dried resin (200 mg) wasthen heated with 2,4-dichloropyrimidine and 250 mg of cesium carbonatein NMP (3 ml) at 80° C. overnight. The resin was washed as before. Onehalf of the resin was heated with 2-(2-aminoethylamino)-5-nitropyridine(180 mg, 1 mmol) in NMP (2 ml) at 125° C. for 66 hours. The resin waswashed as before and then treated with 100% TFA for 0.5 hours. The resinwas filtered off and the filtrate was concentrated in vacuo, thenlyophilized from acetonitrile and water to give the title compound as ayellow solid.

HPLC: 23.46 min (82% purity)

MS: MH⁺=400 C₁₈H₁₈ClN₇O₂=399 g/mol

Example 19 Synthesis ofEthyl-4-{[(3-cyanophenyl)methyl]amino}-2-({2-[(5-nitro(2-pyridyl)amino[ethyl}amino)pyrimidine-5-carboxylate

Bromomethyl Sasrin resin (prepared as in Step 1 of Example 3, 1.0 g) wasreacted with 4-cyanobenzylamine (1.5 ml) in NMP (8 ml) at 80° C. for 4hours. The resin was washed with DMF and dichloromethane and dried invacuo at room temperature. The dried resin (400 mg) was then reactedwith ethyl 2,4-dichloropyrimidine-5-carboxylate (prepared according toV. H. Smith and B. E. Christensen, J. Organic Chem., 20: 829 (1955),which is incorporated herein by reference) (400 mg) and cesium carbonate(400 mg) in NMP (4 ml) at 80° C. overnight. The resin was washed asbefore and dried. The dried resin (200 mg) was then heated with2-(2-aminoethylamino)-5-nitropyridine (180 mg, 1 mmol) in NMP (2 ml) at104° C. for 21 hours. The resin was washed with DMSO, glacial aceticacid, water, DMSO, dichloromethane and then treated with 100% TFA toobtain the title compound.

HPLC: 25.27 min (100% purity)

MS: MH⁺=463 C₂₂H₂₂N₈O₄=462 g/mol

The following additional compounds were prepared according to Resinmethod D using the appropriate amine:

-   (4-{[(3-bromophenyl)methyl]amino}pyrimidin-2-yl)    {2-[(5-nitro(2-pyridyl))amino]ethyl}amine-   [(2,4-dichlorophenyl)methyl][2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]amine-   [(3-methylphenyl)methyl][2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]amine-   [(3,5-dichlorophenyl)methyl][2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]amine-   Ethyl    4-{[(3-bromophenyl)methyl]amino}-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate-   [2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]benzylamine-   [(4-chlorophenyl)methyl][2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]amine-   Ethyl    4-{[(2-chlorophenyl)methyl]amino}-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate-   Ethyl    4-{[(4-cyanophenyl)methyl]amino}-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate

Example 20 Solid Phase Synthesis of Pyridine Compounds Resin Method E

An amino resin (for example Sasrin resin loaded with a primary amine asdescribed in Resin Methods B (Example 3) and D (Example 18)) was reactedwith, e.g., 2,6-dichloro-3-nitropyridine and cesium carbonate in NMP attemperatures within the range of about 25-50° C. for a period within therange of from about 5 hours to about 24 hours. The resin was then washedwith DMF and dichloromethane and heated with a primary amine in NMP attemperatures from 70-100° C. overnight. The resins were washed asdescribed in Example 18, and the pyridine products obtained by treatingthe resin with 20-100% TFA for 0.5-1 hours (preferably with 80-100%TFA).

Example 21 describes the synthesis of compounds of the present inventionpursuant to Resin Method E.

Example 21 Synthesis of{2-[(6-amino-5-nitro(2-pyridyl)amino]ethyl}-{5-nitro-6-[benzylaminol(2-pyridyl)}amine

Step 1: 2-Amino-6-chloro-3-nitropyridine (obtained from2,6-dichloro-3-nitro-pyridine by the method of V. W. von Bebenberg,Chemiker-Zeitung, 103:387 (1979), which is incorporated herein byreference) (2.65 g) was treated at room temperature with ethylenediamine(5 ml). The temperature was gradually raised to 100° C. After 4 h theexcess ethylenediamine was removed by rotary evaporation. The residuewas partitioned between dichloromethane and 2.5 M aqueous sodiumhydroxide. The aqueous layer was further extracted 3 times withdichloromethane. The combined organic layers were concentrated in vacuoto give (2-aminoethyl)(6-amino-5-nitro(2-pyridyl))amine as a canaryyellow solid.

Step 2: Bromomethyl Sasrin resin, prepared according to Step 1 ofExample 3, was heated with benzylamine (2 ml) in NMP (6 ml) at 70° C.for 4 hours. The resin was washed with DMF and dichloromethane and driedin vacuo. The dried resin (100 mg) was heated with2,6-dichloro-3-nitropyridine (190 mg, 1 mmol) and cesium carbonate (100mg) in NMP (2 ml) at 50° C. for 5.5 hours. The resin was then washedwith water, DMF and dichloromethane. The resin was air dried and thenheated with the amine from Step 1 (90 mg) in NMP (2 ml) at 95° C.overnight. The resin was washed with DMSO, acetic acid, water, DMSO,dichloromethane and then treated with 20% TFA to give the titlecompound.

HPLC: 28.47 min (87% purity)

NMR: (300 MHz, 7/1 acetonitrile-d₃/D₂O, 75° C.: 8.0 (2H, two overlappingd), 7.2-7.4 (5H, Ph), 5.9 (2H, 2d overlapping), 4.75 (s, 2H), 3.50-3.65(m, 4H)

The following additional compounds were similarly prepared according toResin Method E by varying the pyridine and primary amine:

-   {2-[(5-nitro(2-pyridyl))amino]ethyl}{5-nitro-6-[benzylamino](2-pyridyl)}amine-   6-{[2-({5-nitro-6-[benzylamino]-2-pyridyl}amino)ethyl]amino}pyridine-3-carbonitrile-   {6-[(2-methoxyethyl)amino]-5-nitro(2-pyridyl)}{2-[(5-nitro(2-pyridyl))amino]ethyl}amine-   (6-{[(2,4-dichlorophenyl)methyl]amino}-5-nitro(2-pyridyl))    {2-[(5-nitro(2-pyridyl))amino]ethyl}amine.

Example 22 Synthesis of4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-5-[benzylamino]pyrimidin-4-yl[benzenecarbonitrile(Resin Method F)

Benzylamine was reacted with bromomethyl Sasrin resin to give abenzylamine substituted resin as in Step 1 of Example 3. This resin (150mg) was shaken with 4-cyanophenacylbromide (130 mg), DMF (2 ml) and2,6-lutidine (200 μl) at room temperature for 6.5 hours. The resin waswashed with DMF and dichloromethane and briefly air dried. It was thenheated with DMFDMA (3 ml) at 80° C. overnight. The resin was then washedwith DMF and dichloromethane and dried in vacuo. The dried resin washeated with amino {2-[(5-nitro(2-pyridyl))amino]ethyl}carboxamidinium4-methylbenzenesulfonate (120 mg) and cesium carbonate (160 mg) in NMP(2 ml) at 90° C. overnight. The resin was washed with DMF, water, DMFand dichloromethane and then treated with 95:5 TFA:water to give thetitle compound.

HPLC: 25.61 min (80% pure)

MS: MH⁺=467 C₂₅H₂₂N₈O₂=466 g/mol

Example 23 Solid Phase Synthesis of Pyrimidines (C₅=carboxyl) ResinMethod G

A mixture of polystyrene Wang resin (Novabiochem, 0.41 mmol/g, 2.2 g,1.21 mmol), a β-ketoester (commercially available from Aldrich orLancaster Chemical, 36.3 mmol) and dimethylaminopyridine (DMAP, 12.1mmol) in toluene (22 ml) was shaken for 16 hours at 90° C. The resin wasfiltered and washed with DCM, DMF, DCM, then dried.

A mixture of the dried resin (100 mg, 0.055 mmol), an aldehyde (0.55mmol), piperidine (0.055 mmol), acetic acid (0.055 mmol) and 3AMolecular Sieves (Aldrich) in DMF (1.0 ml) was shaken for 16 hours atroom temperature. The resin was filtered and washed with DMF and DCM,then dried.

To a mixture of the resulting resin (100 mg, 0.055 mmol) and NaHCO₃ (12mg, 0.138 mmol) was added 0.4 M of the appropriate guanidine in DMF (1.0ml, 0.4 mmol). The mixture was then shaken for 16 hours at 70° C. Theresin was then filtered and washed with DMF, water, methanol, DMF, DCM,and dried.

This resin was treated with 0.1 M DDQ in THF (1.1 ml, 0.11 mmol) for 3hours at room temperature. The resin was filtered and washed with DMF,saturated NaHCO₃ (aq), water, methanol, DMF, DCM, then dried. The resinwas treated with 95% TFA/water for 1 hour at room temperature, thenfiltered and washed with DCM. The filtrate and washings were combinedand evaporated. The residue was dissolved in acetonitrile/water (1:1)then lyophilized.

In all cases, the product pyrimidines were of purity >80% as determinedby HPLC; MS and NMR analysis.

The following compounds were prepared according to Resin Method G usingamino{2-[(5-nitro(2-pyridyl)amino]ethyl}carboxamidinium 4-toluenesulfonate as the guanidine source and the β-ketoester and aldehydeindicated in parentheses:

-   6-(2-Fluorophenyl)-2-({2-[5-nitro(2-pyridyl)amino]ethyl}amino)-4-phenylpyrimidine-5-carboxylic    acid (ethyl 3-(4-fluorophenyl)-3-oxoproprionate and benzaldehyde)-   2-({2-[5-Nitro(2-pyridyl)amino]ethyl}amino)-6-(4-nitrophenyl)-4-phenylpyridine-5-carboxylic    acid (ethyl 3-(4-nitrophenyl)-3-oxoproprionate and benzaldehyde)-   6-Methyl-2-({2-[(5-nitro(2-pyridyl)amino]ethyl}amino)-4-phenylpyrimidine-5-carboxylic    acid (ethyl acetoacetate and benzaldehyde)-   4,6-bis(4-Nitrophenyl)-2-({2-[5-nitro(2-pyridyl)amino]ethyl}amino)pyridine-5-carboxylic    acid (ethyl 3-(4-nitrophenyl)-3-oxoproprionate and    4-nitrobenzaldehyde)-   2-({2-[5-Nitro(2-pyridyl)amino]ethyl}amino)-6-(4-nitrophenyl)-4-(4-pyridyl)pyrimidine-5-carboxylic    acid (ethyl 3-(4-nitrophenyl)-3-oxoproprionate and    4-pyridylcarboxaldehyde)-   4-(4-Methoxyphenyl)-2-({2-[5-nitro(2-pyridyl)amino]ethyl}amino)-6-(4-nitrophenyl)pyrimidine-5-carboxylic    acid (ethyl 3-(4-nitrophenyl)-3-oxoproprionate and    4-methoxybenzaldehyde)-   4-(4-Cyanophenyl)-2-({2-[(5-nitro(2-pyridyl)amino]ethyl}amino)-6-(4-nitrophenyl)pyrimidine-5-carboxylic    acid (ethyl 3-(4-nitrophenyl)-3-oxoproprionate and    4-cyanobenzaldehyde)-   2-({2-[(5-Nitro(2-pyridyl))amino]ethyl}amino)-4-(4-nitrophenyl)pyrimidine-5-carboxylic    acid (ethyl 3-(4-nitrophenyl)-3-oxoproprionate and formaldehyde)-   4,6-bis(4-Cyanophenyl)-2-({2-[(5-nitro(2-pyridyl)amino]ethyl}amino)pyrimidine-5-carboxylic    acid (ethyl 3-(4-cyanophenyl)-3-oxoproprionate and    4-cyanobenzaldehyde)-   4-(4-Cyanophenyl)-2-({2-[(5-nitro(2-pyridyl)amino]ethyl}amino)-6-(3-nitrophenyl)pyrimidine-5-carboxylic    acid (ethyl 3-(4-cyanophenyl)-3-oxoproprionate and    3-nitrobenzaldehyde)-   4-(4-Cyanophenyl)-2-({2-[(5-nitro(2-pyridyl)amino]ethyl}amino)-6-phenylpyrimidine-5-carboxylic    acid (ethyl 3-(4-cyanophenyl)-3-oxproprionate and benzaldehyde)-   4-(3-Cyanophenyl)-2-({2-[(5-nitro(2-pyridyl)amino]ethyl}amino)-6-(4-nitrophenyl)pyrimidine-5-carboxylic    acid (ethyl 3-(4-nitrophenyl)-3-oxoproprionate and    3-cyanobenzaldehyde)-   4-(3-Hydroxyphenyl)-2-({2-(5-nitro(2-pyridyl)amino]ethyl}amino)-6-(4-nitrophenyl)pyrimidine-5-carboxylic    acid (ethyl 3-(4-nitrophenyl)-3-oxoproprionate and    3-hydroxybenzaldehyde)-   2-({2-[(5-Nitro(2-pyridyl))amino]ethyl}amino)-4-(3-nitrophenyl)-6-(4-nitrophenyl)pyrimidine-5-carboxylic    acid (ethyl 3-(4-nitrophenyl)-3-oxoproprionate and    3-nitrobenzaldehyde)-   2-({2-[(5-Nitro(2-pyridyl))amino]ethyl}amino)-6-(4-nitrophenyl)-4-(4-quinolyl)pyrimidine-5-carboxylic    acid (ethyl 3-(4-nitrophenyl)-3-oxoproprionate and    4-quinolinecarboxaldehyde)-   2-({2-[(5-Nitro(2-pyridyl))amino]ethyl}amino)-6-(4-nitrophenyl)-4-[4-(trifluoromethyl)phenyl]pyrimidine-5-carboxylic    acid (ethyl 3-(4-nitrophenyl)-3-oxoproprionate and    4-trifluoromethylbenzaldehyde)-   4-({4-Carboxyphenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-6-(4-nitrophenyl)pyrimidine-5-carboxylic    acid (ethyl 3-(4-nitrophenyl)-3-oxoproprionate and    4-carboxybenzaldehyde)-   4-Cyclohexyl-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-6-(4-nitrophenyl)pyrimidine-5-carboxylic    acid (ethyl 3-(4-nitrophenyl)-3-oxoproprionate and    cyclohexanecarboxaldehyde)-   4-(4-Cyanophenyl)-6-(4-fluorophenyl)-2-({2-[(5-nitro(2-pyridyl)amino]ethyl}amino)pyrimidine-5-carboxylic    acid (ethyl 3-(4-cyanophenyl)-3-oxoproprionate and    4-fluorophenylbenzaldehyde)-   4-(4-Cyanophenyl)-6-(3-furyl)-2-({2-[(5-nitro(2-pyridyl)amino]ethyl}amino)pyrimidine-5-carboxylate    (ethyl 3-(4-cyanophenyl)-3-oxoproprionate and 3-furylcarboxaldehyde)

Example 24 Solid Phase Synthesis of Pyrimidines (C₅=carboxyl, C₄ orC₆=H) Resin Method G

A suspension of resin (Novabiochem, San Diego, USA, 0.51 mmol/g, 100 mg,0.055 mmol) in DMF-dimethylacetal (1 ml) was shaken for 17 hour at roomtemperature. The resin was filtered and washed with DCM and ether, thendried.

To a mixture of the resulting dried resin (100 mg, 0.055 mmol) andNaHCO₃ (12 mg, 0.138 mmol) was added 0.4 M solution of the appropriateguanidine in DMF (1.0 ml, 0.4 mmol). The mixture was shaken for 16 hoursat 70° C. This resin was then filtered and washed successively with DMF,water, MeOH, DMF, DCM, and then dried. The resin was treated with 95%TFA/water for 1 hour at room temperature, then filtered and washed withDCM. The filtrate and washings were combined and evaporated. The residuewas dissolved in acetonitrile:water (1:1 v/v) and lyophilized to give apyrimidine.

The following compounds were prepared according to the above methodusing N-(3-nitropyridine-6-yl)aminoethylguanidine and the appropriateβ-ketoester and aldehyde:

-   4-methyl-2-({2-[(5-nitro(2-pyridyl)amino]ethyl}amino)pyrimidine-5-carboxylic    acid-   2-({2-[(5-nitro(2-pyridyl)amino]ethyl}amino)-4-(4-nitrophenyl)pyrimidine-5-carboxylic    acid-   4-(4-cyanophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-6-(3-nitrophenyl)pyrimidine-5-carboxylic    acid.

Example 25 Solution Phase Synthesis Solution Method A

A carbonyl-containing compound (e.g., β-keto esters, β-keto sulfones,β-keto nitriles, α-nitro ketones, and the like) was dissolved in asuitable organic solvent (usually THF) and treated with a slight excess(1.2-2 equivalents) of DMFDMA. The mixture was heated at 60-80° C. for3-15 hours, most typically 3-5 hours. The reaction mixture was thencooled. When a done on a small scale (0.2-1 mmol) there was no attemptto remove the slight excess of DMFDMA present, rather the cooled mixturewas directly added to a mixture of a guanidine (1 equivalent) and anappropriate base (for example, cesium carbonate or 1.2 equivalents ofsodium ethoxide in 1 ml of ethanol).

The reaction was then heated at 70-80° C. for 12-24 h. At the conclusionof the reaction the vials were cooled, poured into dichloromethane orethyl acetate and washed with saturated aqueous sodium bicarbonatesolution. The organic layer was concentrated in vacuo and the productwas precipitated or crystallized, usually by addition of water toacetonitrile or ethanol solutions of the product. In some caseschromatographic purification was performed, either by semi-preparativeHPLC or by radial chromatography using silica gel plates on aChromatotron (Harrison Research, Palo Alto, Calif.) eluting withmixtures of dichloromethane and methanol. Larger scale reactions wereperformed in round bottom flasks using typical organic chemistryapparatus.

Examples 31, 35-45, and 50-59 describe the synthesis of compounds of thepresent invention pursuant to Solution Method A.

Example 26 Synthesis of Ethyl4-(4-cyanophenyl)-2-{[2-(2-guinolylamino)ethyl]amino}pyrimidine-5-carboxylate

Ethyl 3-(4-cyanophenyl)3-oxopropanoate (64 mg, 0.3 mmol) was heated withDMFDMA (50 μl) and dry THF (1 ml) at 70° C. for 3 hours. The cooledmixture was then added to a suspension ofamino[2-(2-quinolylamino)ethyl]carboxamidinium 4-methylbenzenesulfonate(prepared according to Example 12), (120 mg, 0.3 mmol) in ethanol (2 ml)containing 0.35 mmol of sodium ethoxide. The reaction was then heated at80° C. overnight and then concentrated in vacuo. The residue was takenup in dichloromethane and washed with saturated aqueous sodiumbicarbonate. The organic layer was concentrated in vacuo. The residuewas taken up in acetonitrile. Addition of water gave a precipitate whichwas filtered off and dried to give the title compound.

HPLC: 22.12 min (90% purity)

MS: MH⁺=439 C₂₅H₂₂N₆O₂=438 g/mol

Example 27 Synthesis of Ethyl4-(6-morpholin-4-yl(3-pyridyl))-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate

Step 1: Ethyl 6-chloronicotinate (5.0 g) and morpholine (10 ml) weremixed and then heated to 100° C. In less than 5 minutes at thistemperature, a thick paste formed. Acetonitrile (15 ml) was added andheating was continued overnight at 90° C. The mixture was cooled,diluted with water and extracted with ethyl acetate. The organic layerwas dried and concentrated in vacuo to give ethyl6-morpholin-4-ylpyridine-3-carboxylate as a white solid.

NMR (300 MHz, CDCl₃: 8.80 (s, 1H), 8.05 (d, 1H), 6.60 (d, 1H), 4.35 (q,2H), 3.80 (m, 4H), 3.65 (m, 4H), 1.35 (t, 3H).

The solid was refluxed in a mixture of THF and aqueous potassiumhydroxide for 2 hours. The THF was removed in vacuo and the aqueouslayer was extracted with ethyl acetate. The aqueous layer was thenacidified with acetic acid. A white solid precipitated out and waswashed with water and dried to give6-morpholin-4-ylpyridine-3-carboxylic acid.

NMR (300 MHz, DMSO-d₆) 8.65 (s, 1H), 7.95 (d, 1H), 6.85 (d, 1H), 3.70(m, 4H), 3.60 (m, 4H)

Step 2: The acid described in Step 1 was converted to the β-keto esteras follows. The acid (5.6 g., 27 mmol) in dry THF (100 ml) was treatedat room temperature with oxalyl chloride (40 mmol) followed by severaldrops of DMF. The mixture was then refluxed for 2 hours. The solvent wasremoved in vacuo to give a yellow solid acid chloride. Potassium ethylmalonate (Aldrich Chemical Co., 9.2 g, 54 mmol) and anhydrous magnesiumchloride (6.48 g) were mixed in dry acetonitrile (100 ml). Thentriethylamine (6 ml) was added and the mixture stirred at roomtemperature for 4 hours. An additional 3 ml of triethylamine was added,followed by addition of the acid chloride dissolved in 50 ml of dryacetonitrile. The mixture stirred overnight at room temperature, thenthe solvent was removed in vacuo. The residue was treated with toluene(ca. 200 ml) and then sufficient 25% aqueous HCl was added to dissolvethe residue entirely. The mixture was shaken and organic and aqueouslayers separated. The toluene layer was washed with water. The combinedaqueous layers were then washed twice with toluene. The organic layerswere discarded. The pH of the aqueous layer was adjusted to pH 7 byaddition of solid sodium carbonate. The aqueous layer was then extractedwith toluene. Concentration of the toluene layer in vacuo gave ethyl3-(6-morpholin-4-yl)(3-pyridyl)-3-oxopropanoate as a yellow solid.

Step 3: The β-keto ester from Step 2 (83 mg, 0.3 mmol) was heated withDMFDMA (50 μl) and dry THF (1 ml) at 70° C. for 3 hours. The cooledmixture was then added to a suspension ofamino[2-[(5-nitro(2-pyridyl)amino]ethyl}carboxamidinium4-methylbenzenesulfonate (120 mg, 0.3 mmol) in ethanol (2 ml) containing0.35 mmol of sodium ethoxide. The reaction was then heated at 80° C.overnight and then concentrated in vacuo. The residue was dissolved indichloromethane, then washed with saturated aqueous sodium bicarbonate.This organic layer was then concentrated in vacuo, then redissolved inacetonitrile. Addition of water gave a precipitate which was filteredoff and dried to give the title compound.

HPLC: 18.77 min (98%)

MS: MH⁺=495 C₂₃H₂₆N₈O₅=494 g/mol

Example 28 Synthesis of Ethyl2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(4-cyanophenyl)pyrimidine-5-carboxylate

To a solution of ethyl 3-(4-cyanophenyl)-3-oxopropionate (63 mg, 0.3mmol) in THF (1 ml) was added DMFDMA (50 μl). The solution was heated at70° C. for 3 hours and then added to a mixture ofamino[2-[(5-nitro(2-pyridyl)amino]ethyl}carboxamidinium4-methylbenzenesulfonate (123 mg, 0.3 mmol), ethanol (1 ml) and 1.0 Msodium ethoxide (0.35 ml). This mixture was heated at 80° C. overnight.The reaction was cooled, diluted with dichloromethane and washed withaqueous sodium bicarbonate. The organic layer was concentrated in vacuo,the dissolved in acetonitrile. The product was precipitated with waterto give the title compound.

HPLC: 25.21 min

MS: MH⁺=449 C₂₁H₂₀N₈O₄=448 g/mol

NMR (DMSO-d6): 1.02 (t, 3H), 3.60 (m, 4H), 4.10 (q, 2H), 5.95 (d, 1H),7.60 (d, 2H), 7.85 (d, 2H), 7.90 (d, 1H), 8.80 (s, 1H)

Example 29 Synthesis of ethyl2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(4-morpholin-4-ylphenyl)pyrimidin-5-carboxylate

To a solution of ethyl 3-(4-morpholinophenyl)3-oxopropanoate (70 mg, 0.3mmol) in THF (1 ml) was added DMFDMA (60 μl). The solution was heated at70° C. for 3 hours, then added to a mixture ofamino[2-(6-amino-5-nitro(2-pyridyl)amino]ethyl}carboxamidinium4-methylbenzenesulfonate (123 mg, 0.3 mmol), ethanol (1 ml) and 1.0 Msodium ethoxide (0.35 ml). The mixture was heated at 80° C. overnight,then cooled, diluted with dichloromethane and washed with saturatedaqueous sodium bicarbonate solution. The organic layer was concentratedin vacuo, dissolved in acetonitrile and the product precipitated withwater to give the title compound.

HPLC: 22.37 min. (85% purity)

MS: MH⁺=509 C₂₄H₂₈N₈O₅=508 g/mol

NMR (DMSO-d₆): 1.05 (t, 3H), 3.3 (m, 4H), 3.60 (m, 4H), 3.78 (m, 4H),4.15 (q, 2H), 5.95 (d, 1H), 6.90 (d, 2H), 7.45 (d, 2H), 7.95 9d, 1H),8.60 (s, 1H)

Example 30 Synthesis of Ethyl2-({2-[(6-amino-5-nitro(2-pyridyl)amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidine-5-carboxylate

To a solution of ethyl 3-(2,4-dichlorophenyl)-3-oxopropionate (78 mg,0.3 mmol) in THF (2 ml) was added DMFDMA (70 μl). The solution washeated 3 hours at 70° C., then cooled and added to a suspension ofamino[2-(6-amino-5-nitro(2-pyridyl)amino]ethyl}carboxamidinium4-methylbenzenesulfonate (123 mg, 0.3 mmol), dry ethanol (1 ml) and 1.0M sodium ethoxide (0.35 ml). This mixture was heated at 80° C.overnight. The mixture was cooled, diluted with dichloromethane and thenextracted with saturated aqueous sodium bicarbonate. The organic layerwas concentrated in vacuo and the residual oil was dissolved inacetonitrile. Addition of water gave the title compound as a yellowsolid.

Example 31 Synthesis of Ethyl4-(4-cyanophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate

To a solution of ethyl 3-(4-cyanophenyl)-3-oxopropionate (65 mg, 0.3mmol) in THF (1 ml) was added DMFDMA (50 μl). The solution was heated at70° C. for 3 hours. The solution was then added to a mixture ofamino[2-[(5-nitro(2-pyridyl)amino]ethyl}carboxamidimium4-methylbenzenesulfonate (120 mg, 0.3 mmol), dry ethanol (1 ml) and 1.0M sodium ethoxide (0.35 ml). The mixture was heated at 80° C. overnight.The mixture was cooled, diluted with dichloromethane and washed withsaturated aqueous sodium bicarbonate. The organic layer was concentratedin vacuo, then dissolved in acetonitrile. The solid product wasprecipitated by addition of water to give the title compound.

HPLC: 28.05 min. (95% pure)

Example 32 Synthesis of2-((2-((5-nitro(2-pyridyl)amino)ethyl)amino)-4-(4-cyanophenyl)pyrimidine-5-carboxylate

To a suspension of 1.0 g (2.3 mmol) of ethyl4-(4-cyanophenyl)-2-({2-[(5-nitro(2-pyridyl)amino]ethyl}amino)pyrimidine-5-carboxylate(prepared as described in Example 31) in 1:1 methanol/water was added1.5 mnmol of sodium hydroxide and the solution warmed to 60° C. for 45minutes. During this time the reaction became homogenous. After coolingthe mixture, the pH was adjusted to about 5.0 at which time the desiredacid precipitated from solution. This solid was collected and dried togive 890 mg (2.2 numol, 98% yield) of2-((2-((5-nitro(2-pyridyl)amino)ethyl)amino)-4-(4-cyanophenyl)pyrimidine-5-carboxylateas a light yellow powder.

Example 33 Synthesis of 2-(dimethylamino)ethyl4-(4-cyanophenyl)-2-({2-[(5-nitro(2-pyridyl)amino]ethyl{amino)pyrimidine-5-carboxylate

2-((2-((5-nitro(2-pyridyl)amino)ethyl)amino)-4-(4-cyanophenyl)pyrimidine-5-carboxylate(300 mg, 0.74 mmol) (prepared as described in Example 32) was suspendedin 5 ml of 2-(dimethylamino)ethanol at room temperature.O-Benzotriazole-N,N,N′,N′-tetramethyluronium-hexafluoro-phosphate (HBTU)(Advance Chem Tech, Louisville, Ky.) was then added in one portion andthe mixture stirred for 18 hours at room temperature. The resultingclear solution was poured onto an ice water mixture and extractedthoroughly with ethyl acetate. The aqueous layer was back extracted withethyl acetate 2×. The combined organic layers were then dried withsodium sulfate and concentrated in vacuo. HPLC and NMR analysisindicated that the desired compound, 2-(dimethylamino)ethyl2-((2-((5-nitro(2-pyridyl)amino)ethyl)amino)-4-(4-cyanophenyl)pyrimidine-5-carboxylate,was formed in quantitative yield (>95%).

By substituting an alcohol or amine for the 2-(dimethylamino)ethanolindicated above, the following additional compounds of the presentinvention were similarly synthesized (the alcohol or amine employed isindicated in parentheses):

-   tert-Butyl    4-(4-cyanophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate    (tert-butyl alcohol)-   Methyl    4-(4-cyanophenyl)-2-({2-[((5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate    (methanol)-   Butyl    4-(4-cyanophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate    (n-butanol)-   Phenylmethyl    4-(4-cyanophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate    (benzyl alcohol)-   N-Butyl[4-(4-cyanophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-5-yl]carboxamide    (n-butylamine)-   [4-(4-Cyanophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-5-yl]-N-benzylcarboxamide    (benzylamine)-   [4-(4-Cyanophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-5-yl]-N,N-dimethylcarboxamide    (dimethylamine)-   N-(Cyanomethyl)[4-(4-cyanophenyl)-2-({2-[(5-nitro(2-pyridyl)amino]ethyl}amino)pyrimidin-5-yl]carboxamide    (aminoacetonitrile)-   N-(tert-Butyl)[4-(4-cyanophenyl)-2-({2-[(5-nitro(2-pyridyl)amino]ethyl}amino)pyrimidin-5-yl]carboxamide    (t-butylamine)-   N-[2-(Dimethylamino)ethyl][4-(4-cyanophenyl)-2-({2-[(5-nitro(2-pyridyl)amino]ethyl}amino)pyrimidin-5-yl]carboxamide    38564 (2-(dimethylamino)ethyl amine)-   [4-(4-Cyanophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-5-yl]-N-(2-hydroxyethyl)carboxamide    (2-aminoethanol)-   4-[5-(Morpholin-4-ylcarbonyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]benzenecarbonitrile    (morpholine)-   [4-(4-Cyanophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-2-yl]-N-methylcarboxamide    (methylamino)-   N-(2-Aminoethyl)[4-(4-cyanophenyl)-2-({2-[(5-nitro(2-pyridyl)amino]ethyl}amino)pyrimidin-5-yl]carboxamide    (ethylenediamine)-   4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-5-(piperazinylcarbonyl)pyrimidin-4-yl]benzenecarbonitrile    (piperazine)-   4-(4-Cyanophenyl)-2-({2-[5-nitro(2-pyridyl)amino]ethyl}amino)pyrimidine-5-carboxamide    (ammonia)-   N-(Carbamoylmethyl)[4-(4-cyanophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-5-yl]carboxamide    (glycinamide)-   [4-(4-cyanophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-5-yl]-N-(4-pyridylmethyl)carboxamide-   2-Hydroxyethyl    4-(4-cyanophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate    (ethylene glycol)-   N-(1-Carbamoyl-2-hydroxyethyl)[4-(4-cyanophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-5-yl]carboxamide    (serinamide)

Example 34 Synthesis of4-[5-(3-methyl(1,2,4-oxadiazol-5-yl))-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]benzenecarbonitrile

To a mixture of ethyl4-(4-cyanophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate(0.069 mmol, prepared as described in Example 31) and triethylamine(19.3 μl, 0.14 mmol) in THF (1 ml) was added isobutyl chloroformate(13.4 μl, 0.14 mmol). After stirring at room temperature overnight, theappropriate amidoximine (0.14 mmol) (prepared according to C. D. Bedforeet al., J Med. Chem. 20:2174-2183 (1986), which is incorporated hereinby reference) was added and the mixture stirred at 70° C. for 6 hours.After stirring an additional 72 hours at room temperature, the reactionwas filtered, the solid washed successively with methanol and water, anddried under vacuum to give the desired oxadiazole,4-[5-(3-methyl(1,2,4-oxadiazol-5-yl))-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]benzene-carbonitrile.The following additional compounds were prepared according to thismethod by using the appropriate amidoximine:

-   4-[5-{3-[2-(dimethylamino)ethyl](1,2,4-oxadiazol-5-yl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl)-   (2-{5-[2-({2-[(6-amino-5-nitro(2-pyridyl)amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl](1,2,4-oxadiazol-3-yl)    }ethyl)dimethylamine

Example 35 Synthesis of Ethyl4-(4-morpholin-4-ylphenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate

To a solution of ethyl 3-(4-morpholinophenyl)3-oxopropionate (193 mg,0.7 mmol) in THF (1 ml) was added DMFDMA(140 μl). The solution washeated at 70° C. for 3 hours. This solution was added to a mixture ofamino[2-[(5-nitro(2-pyridyl)amino]ethyl}carboxamidinium4-methylbenzenesulfonate (280 mg, 0.7 mmol), ethanol (1 ml) and 1.0 Msodium ethoxide (0.82 ml). The mixture was heated at 80° C. overnight.The cooled mixture was diluted with dichloromethane and extracted withsaturated aqueous sodium bicarbonate. The organic layer was concentratedin vacuo, then dissolved in acetonitrile. The product was precipitatedas an orange solid (126 mg) by addition of water to give the titlecompound.

HPLC 25.25 min (95% purity)

NMR (DMSO-d6): 1.15 (t, 3H), 3.20 (m, 4H), 3.60 (br. s, 4H), 3.78 (m,4H), 4.05 (q, 2H), 6.59 (d, 1H), 6.95 (d, 2H), 7.40 (d, 2H), 8.0 (m,1H), 8.60 (s, 1H), 8.90 (d, 1H)

MS: MH⁺=494 C₂₄H₂₇N₇O₅=493 g/mol

Example 36 Synthesis of Ethyl4-((4-imidazolylphenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate

To a solution of ethyl 3-[4-(imidazol-1-yl)phenyl]3-oxopropanoate (78mg, 0.3 mmol) (prepared according to I. Sircar et al., J. Med. Chem.,28:1405 (1985), which is incorporated herein by reference) in THF (1 ml)was added DMFDMA (50 μl). The solution was heated at 70° C. for 3 hours.The solution was then added to a mixture ofamino[2-[(5-nitro(2-pyridyl)amino]ethyl}carboxamidinium4-methylbenzenesulfonate (120 mg, 0.3 nmmol), ethanol (1 ml) and 1.0 Msodium ethoxide (0.35 ml). The mixture was heated at 80° C. overnight,cooled, diluted with dichloromethane and washed with saturated aqueoussodium bicarbonate solution. The organic layer was concentrated invacuo, redissolved in acetonitrile and then the product was precipitatedby addition of water as a yellow solid to give the title compound.

HPLC: 18.50 min. (95% purity)

MS: MH⁺=475 C₂₃H₂₂N₈O₄=474 g/mol

NMR (DMSO-d6): 1.05 (t, 3H), 3.62 (br. s, 4H), 4.10 (q, 2H), 6.58 (d,1H), 7.15 (s, 1H), 7.60 (d, 2H), 7.70 (d, 2H), 7.75 (s, 1H), 7.85 (br s,1H), 7.9-8.1 (m, 2H), 8.25 (s, 1H), 8.75 (s, 1H), 8.90 (s, 1H)

Example 37 Synthesis of Ethyl2-({2-[(5-nitro-(2-pyridyl))amino]ethyl}amino)-4-(4-(1,3-oxazol-5-yl)phenyl)pyrimidine-5-carboxylate

Step 1: Methyl 4-formylbenzoate (Aldrich Chemical Co., St. Louis, Mo.)(5.0 g, 30.5 mmol), anhydrous potassium carbonate (4.55 g, 33 mmol) andp-toluenesulfonylmethyl isocyanide (TOSMIC, Aldrich Chemical Co.) (6.83g, 30.5 mmol) were refluxed in methanol (100 ml) for 3.5 hours. Themixture was then concentrated to dryness in vacuo. The residue wasdissolved in ethyl acetate, washed twice with water, dried andconcentrated in vacuo to give methyl 4-(1,3-oxazol-5-yl)benzoate as abeige solid (4.95 g).

(NMR (300 MHz, CDCl₃: 8.10 (d, 2H), 7.98 (s, 1H), 7.75 (d, 2H), 7.48 (s,1H), 3.94 (s, 3H)).

The above ester was heated at reflux for 2 hours in a mixture of 1 Maqueous potassium hydroxide and 50 ml THF. The THF was removed in vacuoand the solution cooled, then acidified with 50% HCl to give4-(l,3-oxazol-5-yl)benzoic acid as a white solid.

NMR (300 MHz, DMSO-d6; 8.52 (s, 1H), 8.05 (d, 2H), 7.82-7.9, m, 3H).

The dried acid above was refluxed in neat thionyl chloride until all ofthe solid had dissolved. The thionyl chloride was removed by rotaryevaporation (with hexane). The crude acid chloride was then dried invacuo briefly. Meanwhile, potassium ethyl malonate (11.1 g, 65 mmol) andanhydrous magnesium chloride (7.7 g, 81 mmol) in dry acetonitrile (150ml) were treated with triethylamine (5.15 ml, 37 mmol). The mixture wasstirred for 3 hours at room temperature, then an additional 1 ml oftriethylamine was added, followed by a solution of the acid chlorideprepared above in dry acetonitrile (50 ml). The reaction was stirredovernight at room temperature. The mixture was concentrated to drynessin vacuo and then partitioned between toluene and 0.25 M aqueous HCl.The organic layer was washed with water, dried and concentrated to givecrude ethyl 3-(4-(1,3-oxazol-5-yl)phenyl)-3-oxopropanoate. The crudeproduct was purified by silica gel chromatography (hexanes/ethylacetate).

Step 2. To a solution of ethyl3-(4-(1,3-oxazol-5-yl)phenyl)-3-oxopropanoate (76 mg, 0.3 mmol) in THF(1 ml) was added DMFDMA (60 μl). The solution was heated at 70° C. for 3hours. This solution was added to a mixture ofamino[2-[(5-nitro(2-pyridyl)amino]ethyl}carboxamidinium4-methylbenzenesulfonate (120 mg, 0.3 mmol), ethanol (1 ml) and 1.0 Msodium ethoxide (0.35 ml). The mixture was heated at 80° C. overnight,cooled, diluted with dichloromethane and washed with saturated sodiumbicarbonate solution. The organic layer was concentrated in vacuo,redissolved in acetonitrile and the product precipitated with water togive the title compound as an orange solid.

HPLC: 26.75 min. (90% purity)

NMR (DMSO-d₆): 1.05 (t, 3H), 3.65 (br. s, 4H), 4.10 (q, 2H), 6.58 (d,1H), 7.58 (d, 2H), 7.70 (s, 1H), 7.75 (d, 2H), 7.82 (br. s, 1H),7.95-8.10 (m, 2H), 8.40 (s, 1H), 8.75 (s, 1H), 8.85 (s, 1H)

Example 38 Synthesis of Ethyl4-(4-(2-furyl)phenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate

Step 1: Ethyl 4-iodobenzoate (2.76 g, 10 mmol) and 2-furylboronic acid(Frontier Scientific, 1.12 g, 10 mmol)) were mixed with bis(triphenylphosphine)palladium dichloride (100 mg) in 1,2-dimethoxyethane(20 ml) and 2 M aqueous sodium carbonate (20 ml). The mixture wasbubbled with argon gas, then heated at 80° C. under argon overnight. Themixture was cooled, diluted with ethyl acetate, then washed with water.The organic layer was dried and concentrated in vacuo to give a crudesolid ester. This material was taken up in a mixture of THF and 1 Maqueous potassium hydroxide and refluxed for 2.5 hours. The THF wasremoved by rotary evaporation and the aqueous layer acidified withacetic acid. Cooling to 4° C. resulted in the precipitation of4-(2-furyl)benzoic acid as a brown solid (1.49 g)

(NMR (300 MHz, DMSO-d6: 8.10 (d, 2H), 7.90 (m, 3H), 7.24 (d, 1H), 6.75(dd, 1H)).

Step 2: The acid from Step 1 was converted to the acid chloride byrefluxing in a mixture of oxalyl chloride (1.3 ml), THF (20 ml) andseveral drops of DMF. Small portions of oxalyl chloride were added untilthe reaction was homogeneous. Reflux continued for 0.5 hours, then thesolvent was removed in vacuo to give the crude acid chloride. Meanwhile,potassium ethyl malonate (2.7 g) was reacted with anhydrous magnesiumchloride (1.9 g) and triethylamine (2.21 ml) in dry acetonitrile (50 ml)at room temperature for 3 hours. Triethylamine (1 ml) was added,followed by addition of a solution of the acid chloride in acetonitrile.The mixture was then stirred overnight at room temperature, thenconcentrated to dryness. The residue was partitioned between toluene and10% aqueous HCl. The organic layer was washed with 10% HCl and water,dried and was then concentrated to give crude ethyl3-(4-(2-furyl)phenyl)-3-oxopropanoate as a solid.

Step 3: The β-keto ester prepared in Step 2 (76 mg, 0.3 mmol) wasdissolved in dry THF (2 ml) and heated with DMFDMA (60 μl) at 70° C. for4 hours. This solution was then added to a mixture ofamino[2-[(5-nitro(2-pyridyl)amino]ethyl}carboxamidinium4-methylbenzenesulfonate (120 mg, 0.3 mmol) and cesium carbonate (160mg) and then heated at 80° C. overnight to give ethyl4-(4-(2-furyl)phenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate.

HPLC: 32.05 min (80% purity)

MS: MH⁺=476 C₂₄H₂₃N₆O₅=475 g/mol

Example 39 Synthesis of Ethyl4-(4-cyanophenyl)-2-({2-[(4-methyl-5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate

Step 1: 2-Chloro-4-methyl-5-nitropyridine (2.0 g, 11.5 mmol) inacetonitrile (10 ml) was added dropwise to ethylenediamine (2.5 ml) inacetonitrile (10 ml). The mixture was stirred overnight at roomtemperature. The solvent was removed by rotary evaporation and theresidue was partitioned between dichloromethane and 2.5 M aqueous sodiumhydroxide. The aqueous layer was further extracted 4 times withdichloromethane. The combined organic layers were washed with asaturated sodium chloride solution, dried and concentrated in vacuo togive (2-aminoethyl)(4-methyl-5-nitro(2-pyridyl))amine as an orange solid(1.74 g).

Step 2: The amine from Step 1 (1.2 g, 6 mmol) was shaken withbenzotriazole carboxamidinium 4-methylbenzenesulfonate (2.0 g, 6 mmol)and DIEA (1.05 ml, 6 mmol) in dry acetonitrile (10 ml) at roomtemperature overnight. Addition of ether resulted in the precipitationof amino{2-[(4-methyl-5-nitro(2-pyridyl))amino]ethyl}carboxamidinium4-methylbenzenesulfonate as a yellow solid.

Step 3: Ethyl 3-(4-cyanophenyl)-3-oxopropanoate (64 mg, 0.3 mmol) in THF(1 ml) and DMFDMA (0.3 mmol) was heated at 70° C. for 3 hours. Thesolution was added to a mixture of the guanidine from Step 2 (123 mg,0.3 mmol), 1.0 M sodium ethoxide in ethanol (0.35 ml) and ethanol (1ml). The mixture was then heated overnight at 80° C., cooled, dilutedwith dichloromethane, then washed with saturated sodium bicarbonatesolution. The organic layer was concentrated in vacuo, redissolved inacetonitrile and the product precipitated with water.

HPLC: 27.63 min (85% pure)

MS: MH⁺=448 C₂₂H₂₁N₇O₄=447 g/mol

Example 40 Synthesis of2-({2-[(5-nitro(2-pyridyl)amino]ethyl}amino)-4-phenylpyrimidine-5-carbonitrile

3-Oxo-3-phenylpropanenitrile (44 mg, 0.3 mmol) in THF (1 ml) and DMFDMA(50 μl) was heated at 70° C. for 3 hours. This solution was added to amixture of amino[2-[(5-nitro(2-pyridyl)amino]ethyl}carboxamidinium4-methylbenzenesulfonate (120 mg, 0.3 mrol), 1.0 M sodium ethoxide inethanol (0.35 ml) and dry ethanol (1 ml) and heated at 80° C. overnight,then concentrated in vacuo. The residue was then taken up indichloromethane and washed with a saturated aqueous sodium bicarbonatesolution. The organic layer was concentrated in vacuo. The residue wastaken up in acetonitrile. Addition of water gave a precipitate which wasfiltered off and dried to give the title compound.

HPLC: 13.87 min (95% pure)

Example 41 Synthesis of{2-[(5-nitro(2-pyridyl))amino]ethyl}-(5-nitro-4-phenylpyrimidin-2-yl)amine

2-Nitro-1-phenylethan-1-one (50 mg, 0.3 mmol) was heated in THF (1 ml)and DMFDMA (50 μl) for 3 h at 70° C. This solution was added to amixture of amino[2-[(5-nitro(2-pyridyl)amino]ethyl}carboxamidinium4-methylbenzenesulfonate (120 mg, 0.3 mmol), 1.0 M sodium ethoxide inethanol (0.35 ml) and dry ethanol (1 ml), heated at 80° C. overnight,then concentrated in vacuo. The residue was taken up in dichloromethaneand washed with saturated aqueous sodium bicarbonate. The organic layerwas concentrated in vacuo. The residue was taken up in acetonitrile.Addition of water gave a precipitate which was filtered off and dried togive the title compound.

HPLC: 15.33 min (100% purity)

MS: MH⁺=382 C₁₇H₁₅N₇O₄=381 g/mol

Example 42 Synthesis of(5-Nitro-4-phenylpyrimidin-2-yl)[2-(2-pyridylamino)ethyl]amine

2-Nitro-1-phenylethan-1-one (50 mg, 0.3 mmol) was heated in THF (1 ml)and DMFDMA (50 μl) for 3 hours at 70° C. This solution was added to amixture of amino[2-(2-pyridyl)amino)ethyl}carboxamidinium4-methylbenzenesulfonate (105 mg, 0.3 mmol), 1.0 M sodium ethoxide inethanol (0.35 ml) and dry ethanol (1 ml), heated at 80° C. overnight,then concentrated in vacuo. The residue was taken up in dichloromethaneand washed with saturated aqueous sodium bicarbonate. The organic layerwas concentrated in vacuo. The residue was taken up in acetonitrile.Addition of water gave a precipitate which was filtered off and dried togive the title compound.

HPLC: 19.66 min (100% purity)

MS: MH⁺=337 C₁₇H₁₆N₆O₂=336 g/mol

Example 43 Synthesis of Ethyl4-(4-cyanophenyl)-2-[(2-{[5-(trifluoromethyl)(2-pyridyl)]amino}ethyl)amino]pyrimidine-5-carboxylate

Step 1: 2-Chloro-5-(trifluoromethyl)pyridine (5.0 g) was heated withethylenediamine (20 ml) at 120° C. overnight. The excess ethylenediaminewas removed by rotary evaporation and the residue was partitionedbetween dichloromethane and 2.5 M aqueous sodium hydroxide. The aqueouslayer was extracted 5 times further with dichloromethane. The combinedorganic layers were washed with a saturated aqueous sodium chloridesolution, dried, then concentrated in vacuo to give(2-aminoethyl)[5-(trifluoromethyl)(2-pyridyl)]amine as an orange oil.

Step 2: The amine from Step 1 (1.1 g, 5.36 numol) was treated withbenzotriazole carboxamidinium 4-methylbenzenesulfonate (1.78 g, 5.36mmol) and DIEA (0.93 ml, 5.36 mmol) in acetonitrile (6 ml) with shakingat room temperature overnight. Addition of ether gaveamino(2-{[5-(trifluoromethyl)(2-pyridyl)]amino}ethyl)carboxamidinium4-methylbenzenesulfonate as a white solid.

Step 3: Ethyl 3-(4-cyanophenyl)-3-oxopropanoate (64 mg, 0.3 mmol) washeated in THF (1 ml) with DMFDMA (50 μl) at 70° C. for 4 hours. Thissolution was added to a mixture of the guanidine from Step 2 (123 mg,0.3 mmol), 1.0 M sodium ethoxide in ethanol (0.35 ml) and dry ethanol (1ml). This mixture was heated at 80° C. overnight, then concentrated invacuo. The residue was taken up in dichloromethane and washed with asaturated aqueous sodium bicarbonate solution. The organic layer wasconcentrated in vacuo. The residue was taken up in acetonitrile.Addition of water gave a precipitate that was filtered off and dried togive the title compound.

HPLC: 24.46 min (85% purity)

MS: MH⁺=457 C₂₂H₁₉N₆O₂F₃=456 g/mol

Example 44 Synthesis of[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine

Step 1. 2,4-Dichlorophenacyl chloride (1.42 g, 6.4 mmol) and imidazole(1.18 g, 16 mmol) were heated in toluene (40 ml) at 75° C. for 2.25hours. The mixture was concentrated to dryness in vacuo. The residue wasdissolved in dichloromethane and washed with 5% aqueous potassiumcarbonate solution and water, dried and concentrated in vacuo. The crudeproduct was purified by passage over a pad of silica gel, eluting with5% methanol in dichloromethane to give1-(2,4-dichlorophenyl)-2-imidazolylethan-1-one as an orange oil.

Step 2: The product of Step 1 (95 mg) was heated with DMFDMA (2 ml) at105° C. for 9 hours. The solvent was removed in vacuo and the residuewas dissolved in dry THF (2 ml) and added to a mixture ofamino[2-[(5-nitro(2-pyridyl)amino]ethyl}carboxamidinium4-methylbenzenesulfonate (100 mg, 0.3 mmol) and cesium carbonate (200mg). The mixture was heated overnight at 80° C., then concentrated invacuo. The residue was taken up in dichloromethane and washed withsaturated aqueous sodium bicarbonate. The organic layer was concentratedin vacuo. The product was purified by radial chromatography on silicagel.

HPLC: 22.48 min (96% purity)

MS: MH⁺=471-473 (cluster, 2 Cl) C₂₀H₁₆Cl₂N₈O₂=471 g/mol

Example 454-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-5-imidazolylpyrimidin-4-yl]benzenecarbonitrile

Step 1: 4-Cyanophenacyl bromide (0.72 g, 3.2 mmol) and imidazole (0.55g, 8 mmol) were heated at 75° C. in toluene (20 ml) for 2.5 hours. Themixture was concentrated to dryness in vacuo. The residue was dissolvedin dichloromethane and washed with a 5% aqueous potassium carbonatesolution and water, dried and concentrated in vacuo to give a pink solid(0.35 g). This method is a variation of the one described in Sakurai etal., 1996, Chem. Pharm. Bull. 44:1510, which is incorporated herein byreference.

Step 2: 1-(4-Cyanophenyl)-2-imidazolylethan-1-one (from Step 1, 63 mg,0.3 mmol) was heated with DMFDMA (2 ml) at 105° C. for 9 hours. Thesolvent was removed in vacuo and the residue was dissolved in dry THF (2ml) and added to a mixture ofamino[2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}carboxamidinium4-methylbenzene-sulfonate (105 mg, 0.3 mmol) and cesium carbonate (200mg). The mixture was heated overnight at 80° C., then concentrated invacuo. The residue was taken up in dichloromethane and washed with asaturated aqueous sodium bicarbonate solution. The organic layer wasconcentrated in vacuo. The residue was crystallized from ethanol/waterto give yellow needles.

HPLC: 17.68 min (100% purity)

MS: MH⁺=443 C₂₁H₁₈N₁₀O₂=442 g/mol

Example 46 Synthesis of[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl)amino]ethyl}amine

A solution of 1-(2,4-dichlorophenyl)-2-imiazol-2-ylethan-1-one (preparedfrom the appropriate acid chloride and 2-methylimidazole according tothe procedure described in Macco et al., J Org. Chem., 20:252 (1985),which is incorporated herein by reference) and DMFDMA (10 ml/mmol ofketone) was stirred at reflux for 12 hours. After concentration of thissolution, the resulting solid was redissolved in DMF (10 ml/mmol).Cs₂CO₃(3 mmol) and (2-(5-nitro(2-pyridyl)amino]ethyl)carboxamidinium4-methylbenzenesulfonate (1.5 mrol) were added, and the mixture stirredfor 8 hours at 100° C. The mixture was cooled, filtered and the filtratediluted with ethyl acetate and washed with saturated aqueous sodiumbicarbonate. Concentration of the organic layers yielded[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl)amino]ethyl}amine.

Example 47 Synthesis of[2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amine

A solution of 1-(2,4-dichlorophenyl)-2-imiazol-2-ylethan-1-one (preparedfrom the appropriate acid chloride and 2-methylimidazole according tothe procedure described in Macco et al., J. Org. Chem., 20:252 (1985),which is incorporated herein by reference) and DMFDMA (10 ml/mmol ofketone) was stirred at reflux for 12 hours. After concentration of thissolution, the resulting solid was redissolved in DMF (10 ml/mmol).Cs₂CO₃(3 mmol) and(2-(6-amino-5-nitro(2-pyridyl)amino)ethyl)carboxamidinium4-methylbenzenesulfonate (1.5 mmol) were added, and the mixture stirredfor 8 hours at 100° C. The mixture was cooled, filtered and the filtratediluted with ethyl acetate and washed with saturated aqueous sodiumbicarbonate. Concentration of the organic layers yielded[2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amine.

Example 48 Synthesis of4-[5-imidazol-2-yl-2-({2-[(5-nitro(2-pyridyl)amino]ethyl}amino)pyrimidin-4-yl]benzenecarbonitrile

A solution of 4-(2-midazol-2-ylacetyl)benzenecarbonitrile (prepared fromthe appropriate acid chloride and 2-methylimidazole according to theprocedure described in Macco et al., J. Org. Chem., 20:252 (1985) andDMFDMA (10 ml/mmol of ketone) was stirred at reflux for 12 hours. Afterconcentration of this solution, the resulting solid was redissolved inDMF (10 ml/mmol). Cs₂CO₃(3 mmol) and(2-(5-nitro(2-pyridyl)amino]ethyl)carboxamidinium4-methylbenzenesulfonate (1.5 minol) were added, and the mixture stirredfor 8 hours at 100° C. The mixture was cooled, filtered and the filtratediluted with ethyl acetate and washed with saturated aqueous sodiumbicarbonate. Concentration of the organic layers yielded4-[5-imidazol-2-yl-2-({2-[(5-nitro(2-pyridyl)amino]ethyl}amino)pyrimidin-4-yl]benzenecarbonitrile.

Example 49 Synthesis of4-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-5-imidazol-2-ylpyrimidin-4-yl]benzenecarbonitrile

A solution of 4-(2-midazol-2-ylacetyl)benzenecarbonitrile (prepared fromthe appropriate acid chloride and 2-methylimidazole according to theprocedure described in Macco et al., J Org. Chem., 20:252 (1985) andDMFDMA (10 ml/mmol of ketone) was stirred at reflux for 12 hours. Afterconcentration of this solution, the resulting solid was redissolved inDMF (10 ml/mmol). Cs₂CO₃(3 mmol) and(2-(6-amino-5-nitro(2-pyridyl)amino)ethyl)carboxamidinium4-methylbenzenesulfonate (1.5 mmol) were added, and the mixture stirredfor 8 hours at 100° C. The mixture was cooled, filtered and the filtratediluted with ethyl acetate and washed with saturated aqueous sodiumbicarbonate. Concentration of the organic layers yielded4-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-5-imidazol-2-ylpyrimidin-4-yl]benzenecarbonitrile.

Example 50 Synthesis of Ethyl2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-4-(4-pyridyl)pyrimidine-5-carboxylate

This compound was prepared from ethyl 3-oxo-3-(4-pyridyl)propanoate andamino[2-[(5-nitro(2-pyridyl)amino]ethyl}carboxamidinium4-methylbenzenesulfonate in accordance with Solution Method A.

HPLC: 17.78 min (90% purity)

NMR(300 MHz, 5/1 acetonitrile-d₃/D₂O): 8.85 (d, 1H), 8.82 (s, 1H), 8.80(d, 2H), 8.01 (dd, 1H), 7.38 (d, 2H), 6.43 (d, 1H), 4.10 (q, 2H),3.60-3.80 (m, 4H), 1.06 (t, 3H).

Example 51 Synthesis of Ethyl4-(3-nitrophenyl)-2-{[2-(2-pyridylamino)ethyl]amino}pyrimidine-5-carboxylate

This compound was prepared from ethyl 3-oxo-3-(3-nitrophenyl)propanoateand amino[2-(2-pyridylamino)ethyl]carboxamidinium4-methylbenzenesulfonate in accordance with Solution Method A.

HPLC: 21.25 min (90% purity)

MS: MH⁺=409 C₂₀H₁₉N₆O₄=408 g/mol

Example 52 Synthesis of Ethyl2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-4-[4-(trifluoromethoxy)phenyl]pyrimidine-5-carboxylate

This compound was prepared from ethyl3-oxo-3-(4-trifluoromethoxyphenyl)propanoate andamino[2-[(5-nitro(2-pyridyl)amino]ethyl}carboxamidinium4-methylbenzenesulfonate in accordance with Solution Method A.

HPLC: 22.50 min (91% purity)

MS: MH⁺=493 C₂₁H₁₉N₆O₅F₃=472 g/mol

Example 53 Synthesis of Ethyl4-(3,4-difluorophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate

This compound was prepared from ethyl3-oxo-3-(3,4-difluorophenyl)propanoate andamino[2-[(5-nitro(2-pyridyl)amino]ethyl}carboxamidinium4-methylbenzenesulfonate in accordance with Solution Method A.

HPLC: 17.96 min (100% purity)

MS: MH⁺=445 C₂₀H₁₈N₆O₄F₂=444 g/mol

Example 54 Synthesis of Ethyl4-[4-(methylsulfonyl)phenyl]-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate

This compound was make from ethyl3-(4-methylsulfonylphenyl)-3-oxopropanoate andamino[2-[(5-nitro(2-pyridyl)amino]ethyl}carboxamidinium4-methylbenzenesulfonate in accordance with Solution Method A.

HPLC: 11.21 min (100% purity)

MS: MH⁺=487 C₂₁H₂₂N₆O₆S=486 g/mol

Example 55 Synthesis of Ethyl4-(4-methylthiophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate

This compound was prepared from ethyl3-(4-methylthiophenyl)-3-oxopropanoate andamino[2-[(5-nitro(2-pyridyl)amino]ethyl}carboxamidinium4-methylbenzenesulfonate in accordance with Solution Method A.

HPLC: 17.26 min (92% purity)

MS: MH⁺=455 C₂₁H₂₂N₆O₄S=454 g/mol

Example 56 Synthesis of Ethyl4-[4-(dimethylamino)phenyl]-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate

This compound was prepared from ethyl3-(4-dimethylaminophenyl)-3-oxopropanoate andamino[2-[(5-nitro(2-pyridyl)amino]ethyl}carboxamidinium4-methyl-benzenesulfonate in accordance with Solution Method A.

HPLC: 9.0 min (90% purity)

MS: MH⁺=452 C₂₂H₂₅N₇O₄=451 g/mol

Example 57 Synthesis of Ethyl2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(4-cyanophenyl)pyrimidine-5-carboxylate

This compound was prepared from ethyl 3-(4-cyanophenyl)-3-oxopropanoateand amino{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}carboxamidinium4-methylbenzene-sulfonate in accordance with Solution Method A.

HPLC: 25.21 min (83% purity)

MS: MH⁺=449 C₂₁H₂₀N₈O₄=448g/mol

Example 58 Synthesis of Ethyl4-(4-imidazolylphenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate

This compound was prepared from ethyl3-[4-(1-imidazolyl)phenyl]-3-oxopropanoate andamino[2-[(5-nitro(2-pyridyl)amino]ethyl}carboxamidinium4-methylbenzenesulfonate in accordance with Solution Method A.

HPLC: 18.50 min (91% purity)

MS: MH⁺=475 C₂₃H₂₂N₈O₄=474 g/mol

Example 59 Synthesis of Ethyl4-(4-ethylphenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate

This compound was prepared from ethyl 3-(4-ethylphenyl)-3-oxopropanoateand amino[2-[(5-nitro(2-pyridyl)amino]ethyl}carboxamidinium4-methylbenzenesulfonate in accordance with Solution Method A.

HPLC: 32.45 min (95% purity)

MS: MH⁺=437 C₂₂H₂₄N₆O₄=436 g/mol

The following additional compounds were prepared according to SolutionMethod A using the appropriate carbonyl containing compound andguanidine.

-   Ethyl    4-(2-furyl)-2-[(2-(2-pyridyl)ethyl)amino]pyrimidine-5-carboxylate-   Ethyl    4-(3-nitrophenyl)-2-[(2-(2-pyridyl)ethyl)amino]pyrimidine-5-carboxylate-   Ethyl    4-(4-fluorophenyl)-2-[(2-(2-pyridyl)ethyl)amino]pyrimidine-5-carboxylate-   Ethyl    4-(4-methoxyphenyl)-2-[(2-(2-pyridyl)ethyl)amino]pyrimidine-5-carboxylate-   Ethyl    4-(4-cyanophenyl)-2-[(2-(2-pyridyl)ethyl)amino]pyrimidine-5-carboxylate-   2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-4-(4-nitrophenyl)pyrimidine-5-carboxylic    acid-   Ethyl    4-(4-fluorophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate-   Ethyl    2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-4-(3-quinolyl)pyrimidine-5-carboxylate-   Ethyl    2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-4-(3-nitrophenyl)pyrimidine-5-carboxylate-   Methyl    2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-4-(3-pyridyl)pyrimidine-5-carboxylate-   Ethyl    2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-4-(4-nitrophenyl)pyrimidine-5-carboxylate-   Ethyl 4-[3    ,5-bis(trifluoromethyl)phenyl]-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate-   Ethyl    2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-4-[4-(trifluoromethyl)phenyl]pyrimidine-5-carboxylate-   Ethyl    2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-4-[3-(trifluoromethyl)phenyl]pyrimidine-5-carboxylate-   Ethyl    4-(5-bromo(3-pyridyl))-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate-   Ethyl    4-(2,4-difluorophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate-   Ethyl    4-(2,4-dichlorophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate-   Ethyl    4-(4-cyanophenyl)-2-{[2-(pyrimidin-2-ylamino)ethyl]amino}pyrimidine-5-carboxylate-   Ethyl    4-(4-methoxyphenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate-   Ethyl    4-(3-cyanophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate-   Ethyl    4-(4-cyanophenyl)-2-({2-[(4-nitrophenyl)amino]ethyl}amino)pyrimidine-5-carboxylate-   Ethyl    4-(3-fluorophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate-   Ethyl    4-(3,5-dichlorophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate-   4-[5-(methylsulfonyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]benzenecarbonitrile-   Ethyl    2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-4-(4-sulfamoylphenyl)pyrimidine-5-carboxylate-   Ethyl    4-(4-chlorophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate-   Ethyl    4-(4-bromophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate-   Ethyl    4-naphthyl-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate-   Ethyl    2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-4-(4-phenylphenyl)pyrimidine-5-carboxylate-   Ethyl 4-(2H-benzo[3,4-d]1    ,3-dioxolen-5-yl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate-   Ethyl    4-(4-butoxyphenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate-   6-[(2-{[4-(4-cyanophenyl)-5-(ethoxycarbonyl)pyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carboxylic    acid-   tert-Butyl    4-(4-cyanophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate-   tert-Butyl    6-[(2-{[4-(4-cyanophenyl)-5-(ethoxycarbonyl)pyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carboxylate-   Methyl    4-(4-cyanophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate-   Methyl    6-[(2-{[4-(4-cyanophenyl)-5-(ethoxycarbonyl)pyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carboxylate-   Ethyl    4-(4-cyanophenyl)-2-[(2-{[5-(morpholin-4-ylcarbonyl)(2-pyridyl)]amino}ethyl)amino]pyrimidine-5-carboxylate-   Ethyl    4-(4-cyanophenyl)-2-[(2-{[5-(N-ethylcarbamoyl)(2-pyridyl)]amino}ethyl)amino]pyrimidine-5-carboxylate-   4-[5-nitro-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]benzenecarbonitrile-   Ethyl    4-(4-cyanophenyl)-2-{[2-({5-nitro-6-[benzylamino](2-pyridyl)}amino)ethyl]amino}pyrimidine-5-carboxylate-   Ethyl    4-[4-(4-methylpiperazinyl)phenyl]-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate-   Ethyl    2-({2-[(5-cyano(2-pyridyl))amino]ethyl}amino)-4-(4-cyanophenyl)pyrimidine-5-carboxylate-   Ethyl    4-(4-cyanophenyl)-2-[(2-{[6-(methylamino)-5-nitro(2-pyridyl)]amino}ethyl)amino]pyrimidine-5-carboxylate-   Ethyl    2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-4-(4-(1,3-oxazol-5-yl)phenyl)pyrimidine-5-carboxylate-   Ethyl    2-({2-[(4-amino-5-nitropyrimidin-2-yl)amino]ethyl}amino)-4-(4-cyanophenyl)pyrimidine-5-carboxylate-   Ethyl    2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(4-(1,3-oxazol-5-yl)phenyl)pyrimidine-5-carboxylate-   Ethyl    4-[4-(methylethyl)phenyl]-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate-   Ethyl    4-[4-(tert-butyl)phenyl]-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate-   Ethyl    4-(3,4-dichlorophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate-   Ethyl    4-(3,4-dimethoxyphenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate-   Ethyl    4-[4-(diethylamino)phenyl]-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate-   2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4,6-trichlorophenyl)pyrimidine-5-carboxylic    acid-   Ethyl    4-(4-methylphenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate-   Ethyl    4-(2-naphthyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate-   Ethyl    4-(3,4-dimethylphenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate-   Ethyl    2-({2-[(4-amino-5-cyanopyrimidin-2-yl)amino]ethyl}amino)-4-(4-cyanophenyl)pyrimidine-5-carboxylate-   4-(2-methoxyphenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylic    acid-   Ethyl    4-(4-cyanophenyl)-2-{[2-(3-methoxyphenyl)ethyl]amino}pyrimidine-5-carboxylate-   2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(3,4-dichlorophenyl)pyrimidine-5-carbonitrile-   4-(3,4-dichlorophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carbonitrile-   Ethyl    2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-4-(4-(1,2,4-triazol-4-yl)phenyl)pyrimidine-5-carboxylate-   2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidine-5-carbonitrile-   4-(2,4-dichlorophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carbonitrile-   2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidine-5-carboxylic    acid-   Ethyl    2-({2-[(5-amino(2-pyridyl))amino]ethyl}amino)-4-(4-cyanophenyl)pyrimidine-5-carboxylate

Example 60 Solution Phase Synthesis Solution Method B

A ketone with a CH₂ or CH₃ group adjacent to the carbonyl group washeated in neat N,N-dimethylformamide dimethyl acetal (DMFDMA) at 90-110°C. for 5 to 24 hours, usually 8 to 14 hours. The excess DMFDMA was thenremoved by rotary evaporation to give the intermediate enaminoketone asan oil or solid. This intermediate could be crystallized if desired, butwas usually used in crude form in the next reaction step. Theenaminoketone was dissolved in an appropriate solent such as THF,ethanol, isopropanol or for syntheses where a higher reactiontemperature was desired, in NMP (ca. 1-2 ml of solvent for 0.3-1 mmol ofstarting ketone).

This solution was then added to a mixture of a guanidine (1 equivalent)and a suitable base such as sodium ethoxide (freshly prepared), cesiumcarbonate or powdered sodium hydroxide. The usual combinations werecesium carbonate in THF or sodium ethoxide in ethanol or sodiumhydroxide in isopropanol or cesium carbonate in NMP, although other baseand/or solvent combinations can be used. The reaction was then heated at80-125° C. (depending on the boiling point of the solvent) for 12 to 66hours.

Small scale (i.e., 0.2-1 mmol) reactions were conducted in screw capvials. The vials were placed into predrilled thermostated aluminumblocks (Digi-Block, Laboratory Devices, Holliston, Mass.) and shaken ona gyrotary shaker (Lab-Line Model G-2). After completion of thereaction, the vials were cooled, and their contents poured intodichloromethane or ethyl acetate, then washed with saturated aqueoussodium bicarbonate solution. The organic layer was concentrated in vacuoand the product was precipitated or crystallized, usually by addition ofwater to acetonitrile or ethanol solutions of the product. In some caseschromatographic purification was performed, either by semi-preparativeHPLC or by radial chromatography using silica gel plates on aChromatotron (Harrison Research, Palo Alto, Calif.) eluting withmixtures of dichloromethane and methanol. Larger scale reactions wereperformed in round bottom flasks using typical organic chemistryapparatus.

Examples 61-66 describe the synthesis of compounds prepared pursuant toSolution Method B.

Example 61 Synthesis of[2-(2-pyridylamino)ethyl](4-(3-pyridyl)pyrimidin-2-yl)amine

3-Acetylpyridine (0.5 mmol) was heated with DMFDMA (300 μl) at 90° C.for 8.5 hours. The solvent was removed by rotary evaporation. Theresidue was dissolved in isopropanol (2 ml) and added to 170 mg (0.5mmol) of amino[2-(2-pyridylamino)ethyl]carboxamidinium4-methylbenzenesulfonate and powdered sodium hydroxide (70 mg). Themixture was heated at 85° C. overnight, then concentrated in vacuo. Theresidue was taken up in dichloromethane and washed with a saturatedaqueous sodium bicarbonate solution. The organic layer was concentratedin vacuo. The residue was taken up in acetonitrile. Addition of watergave a precipitate that was filtered off and dried to give the titlecompound.

HPLC: 9.9 min(100% purity)

NMR (300 MHz, 5/1 acetonitrile-d₃/D₂O, 75° C.): 9.20 (s, 1H), 8.65 (d,1H), 8.2-8.4 (m, 2H), 7.94 (d, 1H), 7.50 (dd, 1H), 7.38 (t, 1H), 7.10(d, 1H), 6.50 (m, 2H), 3.70 (t, 2H), 3.50 (t, 2H)

Example 62 Synthesis of(5-Ethyl-4-phenylpyrimid-2-yl)[2-(2-pyridylamino)ethyl]amine

Butyrophenone (0.5 mmol) was heated with DMFDMA (300 μl) at 90° C. for8.5 hours. The solvent was removed by rotary evaporation. The residuewas dissolved in isopropanol (2 ml) and added to 170 mg (0.5 mmol) ofamino[2-(2-pyridylamino)ethyl]carboxamidinium 4-methylbenzenesulfonateand powdered sodium hydroxide (70 mg). The mixture was heated at 90° C.overnight, then concentrated in vacuo. The residue was taken up indichloromethane and washed with saturated aqueous sodium bicarbonate.The organic layer was concentrated in vacuo. The residue was taken up inacetonitrile. Addition of water gave a precipitate which was filteredoff and dried to give the title compound.

HPLC: 17.46 min (98% purity)

MS: MH⁺=320 C₁₈H₂₁N₅=319 g/mol

Example 63 Synthesis of[2-(2,5-dimethoxyphenyl)ethyl](4-(3-pyridyl)pyrimidin-2-yl)amine

Step 1: 2,5-Dimethoxyphenethylamine (1.08 g, 6 mmol) was shaken withbenzotriazole carboxamidinium 4-methylbenzenesulfonate (2.0 g, 6 mmol)and DIEA (1.05 ml, 6 mmol) in dry acetonitrile (10 ml) at roomtemperature overnight. Addition of ether resulted in the precipitationof amino[2-(2,5-dimethoxyphenyl) ethyl]carboxamidinium4-methylbenzenesulfonate as a white solid.

Step 2: 3-Acetylpyridine (37 mg, 0.3 imol) was heated at 100° C. inDMFDMA (1 ml) for 8 hours. The solvent was removed by rotary evaporationand the residue was dissolved in dry THF (2 ml) and added to a mixtureof cesium carbonate (160 mg) and 120 mg (0.3 mmol) of the guanidineprepared in Step 1. The mixture was then heated at 80° C. overnight,then concentrated in vacuo. The residue was then taken up indichloromethane and washed with a saturated aqueous sodium bicarbonatesolution. The organic layer was concentrated in vacuo. The residue wastaken up in ethanol (2 ml). Addition of water gave a precipitate whichwas filtered off and dried to give the title compound.

HPLC: 18.03 min (100% purity)

MS: MH⁺=337 C₁₉H₂₀N₄O₂=336 g/mol

Example 64 Synthesis of[4-(4-Morpholin-4-ylphenyl)pyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine

4-Morpholinoacetophenone (0.633 g, 2.5 mmol) was heated at 100° C. in 4ml of DMFDMA for 9 hours. The mixture was concentrated to a viscous oilby rotary evaporation. The oil was redissolved in isopropanol (10 ml)and treated with amino[2-[(5-nitro(2-pyridyl)amino]ethyl}carboxamidinium4-methylbenzenesulfonate (1.0 g, 2.5 inmol) and powdered sodiumhydroxide (200 mg). This mixture was heated at 80° C. overnight. Thecooled mixture was diluted with dichloromethane and washed withsaturated aqueous sodium bicarbonate solution. The organic layer wasconcentrated in vacuo, the dissolved in acetonitrile. The product wasprecipitated by addition of water. The brown solid was recrystalizedfrom isopropanol to give the title compound.

M.P. 223-225° C. (with decomposition)

Elemental Analysis; C₂₁H₂₃N₇O₃. 0.7 H₂O requires C, 58.10; H, 5.66; N,22.59; found C, 58.02; H, 5.30N; 22.39;

HPLC: 20.85 min (100% purity)

MS: MH⁺=422 g/mol (FW=421)

NMR (DMSO-d₆): 3.30 (m, 4H), 3.60 (m, 4H), 3.75 (m, 4H), 6.58 (d, 1H),6.95 (m, 3H), 8.00 (d, 2H), 8.10 (d, 1H), 8.25 (d, 1H), 8.90 (s, 1H)

Example 65 Synthesis of[4-(2,4-dichlorophenyl)-5-ethylpyrimid-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine

Step 1: A mixture of 2,4-dichlorobenzoyl chloride (4.5 g) and copper (I)iodide (200 mg) in dry THF (30 ml) was cooled to −20° C. under argon.Then a solution of n-propyl magnesium chloride (2 M in ether, 11.0 ml)was added dropwise. Ten minutes after addition was complete, the coolingbath was removed and the mixture stirred for 1 hour. Water was carefullyadded, followed by extraction with toluene. The toluene layer was washedwith dilute HCl, water, saturated sodium bicarbonate solution, dried andconcentrated in vacuo to give 1-(2,4-dichlorophenyl)butan-1-one (4.0 g).

Step 2: The ketone from Step 1 (108 mg, 0.5 mmol) was heated at 95° C.overnight with DMFDMA (1.5 ml). The solvent was removed in vacuo and theresidue was dissolved in dry ethanol (2 ml) and added to a mixture ofamino[2-[(5-nitro(2-pyridyl)amino]ethyl}carboxamidinium4-methylbenzenesulfonate (200 mg), 1.0 M sodium ethoxide (0.6 ml) anddry ethanol (2 ml). This mixture was heated at 85° C. overnight, thenconcentrated in vacuo, redissolved in dichloromethane and washed withsaturated sodium bicarbonate solution. The organic layer wasconcentrated in vacuo. The residue was purified by chromatography onsilica gel using 10% methanol in dichloromethane to give an oilyproduct. Lyophilization from acetonitrile/water gave the title compoundas a solid.

HPLC: 29.56 (85% purity)

MS: MH⁺=433 Cl₁₉H₁₈N₆Cl₂O₂=432 g/mol

Example 66 Synthesis of[4-(4-imidazolylphenyl)pyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine

4-(1-Imidazolyl)acetophenone (57 mg, 0.3 mmol) was heated with DMFDMA (1ml) for 8 hours at 105° C. The solvent was removed in vacuo and theresidue was dissolved in dry THF (2 ml) and added to a mixture ofamino[2-[(5-nitro(2-pyridyl)amino]ethyl}carboxamidinium4-methylbenzenesulfonate (120 mg, 0.3 mmol) and cesium carbonate (200mg) and heated overnight at 80° C., then concentrated in vacuo,redissolved in dichloromethane and washed with a saturated sodiumbicarbonate solution. The organic layer was concentrated in vacuo. Theresidue was purified by crystallization to give the title compound.

HPLC: 15.17 min (100% purity)

NMR (300 Mhz, DMSO-d₆): 8.90 (s, 1H), 8.38 (d, 1H), 8.30 (s, 1H), 8.22(d, 2H), 8.05 (d, 1H), 7.75 (d, 2H), 7.20 (s, 1H), 7.15 (d, 1H), 6.58(d, 1H), 3.60 (m, 4H).

The following additional compounds were similarly prepared according toSolution Method B by varying the ketone and guanidine used:

-   (4-phenylpyrimidin-2-yl)(2-(2-pyridyl)ethyl)amine-   4-(2-{[2-(2-pyridylamino)ethyl]amino}pyrimidin-4-yl)benzenecarbonitrile-   (4-phenylpyrimidin-2-yl) [2-(2-pyridylamino)ethyl]amine-   4-{2-[(2-(2-pyridyl)ethyl)amino]pyrimidin-4-yl}benzenecarbonitrile-   [4-(4-nitrophenyl)pyrimidin-2-yl][2-(2-pyridylamino)ethyl]amine-   [4-(4-imidazolylphenyl)pyrimidin-2-yl][2-(2-pyridylamino)ethyl]amine-   [4-(3    ,4-difluorophenyl)pyrimidin-2-yl][2-(2-pyridylamino)ethyl]amine-   [2-(2-pyridylamino)ethyl]{4-[4-(trifluoromethoxy)phenyl]pyrimidin-2-yl}amine-   [4-(2,4-dichlorophenyl)pyrimidin-2-yl][2-(2-pyridylamino)ethyl]amine-   [4-(4-chlorophenyl)-5-methylpyrimidin-2-yl][2-(2-pyridylamino)ethyl]amine-   [4-(4-methyl-1-phenylpyrazol-3-yl)pyrimidin-2-yl][2-(2-pyridylamino)ethyl]amine-   3-[2-({2-[(5-nitro-2-pyridyl)amino]ethyl}amino)pyrimidin-4-yl]benzenecarbonitrile-   [4-(2,4-dimethyl(1,3-thiazol-5-yl))pyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine-   {2-[(5-nitro(2-pyridyl))amino]ethyl}(4-pyrazin-2-ylpyrimidin-2-yl)amine-   [4-phenyl-5-benzylpyrimidin-2-yl][2-(2-pyridylamino)ethyl]amine-   4-[2-({2-[(5-nitro-2-pyridyl)amino]ethyl}amino)pyrimidin-4-yl]benzenesulfonamnide-   {4-[4-(4,5-dichloroimidazol-2-yl)phenyl]pyrimidin-2-yl}{2-[(5-nitro(2-pyridyl))amino]ethyl}amine-   4-(2-{[2-(2,5-dimethoxyphenyl)ethyl]amino}pyrimidin-4-yl)benzenecarbonitrile-   [2-(2,5-dimethoxyphenyl)ethyl](4-(3-pyridyl)pyrimidin-2-yl)amine-   [4-(4-benzimidazolylphenyl)pyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine-   4-[5-imidazolyl-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]benzenecarbonitrile-   4-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-5-imidazolylpyrimidin-4-yl]benzenecarbonitrile-   [4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine-   {2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl]amine-   [4-(2,4-dimethylphenyl)-5-imidazolylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine-   ethyl    4-[2-({2-[(5-nitro-2-pyridyl)amino]ethyl}amino)pyrimidin-4-yl]benzoate-   4-(2-{[3-(4-phenylimidazolyl)propyl]amino}pyrimidin-4-yl)benzenecarbonitrile-   (3-benzimidazolylpropyl)[4-(4-imidazolylphenyl)pyrimidin-2-yl]amine-   N-{4-[2-({2-[(5-nitro-2-pyridyl)amino]ethyl}amino)pyrimidin-4-yl]phenyl}acetamide

Example 67 Synthesis of[5-(4-(Fluorophenyl)pyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amineSolution Method C

Step 1: Dry DMF (22 ml) was cooled to 0° C. under argon. Phosphorousoxychloride (9.2 g) was added dropwise to the cooled DMF. The mixturewas removed from the cooling bath and stirring continued for 1 hour.Then, 4-fluorophenylacetic acid (3.08 g, 20 mnmol) was added as a solidand the mixture was heated at 85° C. for 6 hours. After the mixture wascooled to room temperature it was poured onto approximately 100 g of icewith stirring. A solution of sodium perchlorate monohydrate (3.66 g) inwater (10 ml) was added. The precipitated solid was filtered, washedwith water and dried in vacuo to give[(2-E,Z)-3-(dimethylamino)-2-(4-fluorophenyl)prop-2-enylidene]dimethyl-ammoniumperchlorate. This procedure is described in Church et al., J. Org.Chem., 60:3750 (1995), which is incorporated herein by reference.

Step 2: The vinylogous amidinium salt obtained in Step 1 (100 mg, 0.3mmol) was treated with dry ethanol (2 ml) andamino{2-[(5-nitro(2-pyridyl)amino]ethyl}carboxamidinium4-methylbenzenesulfonate (180 mg, 0.45 mmol). Then, 0.45 ml of a 1.0 Msolution of sodium ethoxide in ethanol was added and the mixture wasshaken 0.5 hours at room temperature. Another 0.3 ml of sodium ethoxidesolution was added, followed by heating at 70° C. for 2 hours. Thesolvent was removed in vacuo. The residue was partitioned betweendichloromethane and water. The organic layer was dried and concentratedin vacuo, then the resulting residue was dissolved in acetonitrile.Addition of water to the residue/acetonitrile mixture caused the titlecompound to precipitate as an orange solid.

HPLC: 18.49 min (80% purity)

NMR (300 MHz, DMSO-d₆: 8.90 (d, 1H), 8.60 (s, 2H), 8.12 (dd, 1H), 7.65(m, 2H), 7.24 (m, 2H), 6.60 (d, 1H), 3.58 (m, 4H)

Example 68 Synthesis of Ethyl4-[(2,4-dichlorophenyl)amino]-2-({2-[(5-nitro(2-pyridyl)amino]ethyl}amino)pyrimidine-5-carboxylateSolution Method D

Step 1: Ethyl 2,4-dichloropyrimidine-5-carboxylate (0.49 g, 2 mmol) and2,4-dichloroaniline (0.33 g, 2 mmol) and DIEA (0.35 ml, 2 mmol) inacetonitrile (6 ml) were heated at 80° C. for 36 hours. The mixture wascooled and the crystalline product, ethyl4-[(2,4-dichlorophenyl)amino]-2-chloropyrimidine-5-carboxylate, 0.54 gwas filtered off.

NMR (300 MHz, CDCl₃): 8.90 (s, 1H), 8.44 (d, 1H), 7.45 (d, 1H), 7.32(dd, 1H), 4.45 (q, 2H), 1.45 (t, 3H)].

Step 2: The pyrimidine from Step 1 (69 mg, 0.2 mmol) was heated withDIEA (100 μL), and (2-aminoethyl)(5-nitro(2-pyridyl))amine (36 mg, 0.2mmol) in NMP (3 ml) at 105° C. for 14 hours. The reaction was cooled,poured into water and extracted with ethyl acetate. The organic layerwas separated, washed with water, dried, then concentrated in vacuo. Thecrude product was purified by radial chromatography on silica gel,followed by crystallization from a mixture of acetonitrile, methanol andwater to give colorless crystals.

HPLC: 30.32 min (>95% purity)

MS: MH⁺=492-494 (cluster) C₂₀H₁₉N₇O₄Cl₂=492 g/mol

Example 69 Synthesis of tert-Butyl6-[(2-{[4-(4-cyanophenyl)-5-ethoxycarbonyl)pyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carboxylate

Step 1: 6-Chloro-pyridine-3-carboxylic acid (5.6 g, 36 mmol) was treatedwith 1,1′-carbonyldiimidazole (6.93 g, 42 mmol) in DMF (40 ml) at 40° C.for 1 hour. Then, t-butanol (9.5 ml, 0.11 mol) and1,8-Diazabicyclo[5.4.0]undec-7-ene (DBU) (5.38 ml, 36 mmol) were addedand heating continued overnight. The mixture was cooled to roomtemperature and diluted with ether (300 ml). The mixture was extractedonce with water. The aqueous layer was back-extracted twice withdichloromethane. The combined organic layers were washed with asaturated aqueous citric acid solution, dried and concentrated in vacuoto give a cream colored solid (7.07 g).

(NMR (300 MHz, CDCl₃): 8.92 (d, 1H), 8.20 (dd, 1H), 7.40 (d, 1H), 1.60(s, 9H)).

The tert-butyl 6-chloropyridine-3-carboxylate was heated withethylenediamine (20 ml) at 80° C. overnight. The solvent was removed invacuo. The residue was partitioned between dichloromethane and 2.5 Maqueous sodium hydroxide solution. The aqueous layer was extracted afurther three times with dichloromethane. The combined organic layerswere washed with water, dried and concentrated in vacuo to givetert-butyl 6-[(2-aminoethyl)amino]pyridine-3-carboxylate.

NMR (300 MHz, CDCl₃): 8.70 (s, 1H), 7.95 (d, 1H), 6.40 (d, 1H), 3.42 (m,2H), 2.96 (m, 2H), 1.70 (s, 9H)

Step 2: t-Butyl 6-[(2-aminoethyl)amino]pyridine-3-carboxylate (1.42 g, 6mmol), benzotriazole carboxamidinium 4-methylbenzenesulfonate (2.0 g, 6mmol) and DIEA (1.05 ml, 6 mmol) were shaken in a mixture of dryacetonitrile (10 ml) and DMF (2 ml) overnight. Ether was added, followedby cooling 4 days at 4° C. The solid was filtered off and dried in vacuoto give tert-butyl6-{[(2-(amidinoammonium)ethyl]amino}pyridine-3-carboxylate4-methylbenzenesulfonate (1.87 g).

NMR (300 MHz, DMSO-d₆: 8.55 (br s, 1H), 7.80 (d, 1H), 7.55 (d, 2H), 7.10(d, 2H), 6.50 (d, 1H), 3.50 (m, 2H), 3.30 (m, 2H), 2.30 (s, 3H), 1.52(s, 9H).

Step 3: Ethyl 3-(4-cyanophenyl)-3-oxopropanoate (217 mg, 1.0 mmol) washeated with DMFDMA (200 μl) in dry THF (2 ml) at 70° C. for 5.5 hours.To the cooled solution was added tert-butyl6-{[(2-(amidinoammonium)ethyl]amino}pyridine-3-carboxylate4-methylbenzenesulfonate (451 mg, 1.0 mmol) along with dry ethanol (4ml) and 1.0 M sodium ethoxide in ethanol (1.2 ml). The mixture washeated at 80° C. overnight. The solvents were removed in vacuo. Theresidue was partitioned between dichloromethane and the saturatedaqueous sodium bicarbonate solution. The organic layer was concentratedin vacuo. The residue was dissolved in acetonitrile. Addition of watergave the title compound as a solid (230 mg).

HPLC: 25.90 min (80% purity)

MS: MH⁺=489 C₂₆H₂₈N₆O₄=488 g/mol

Example 70 Synthesis of6-[(2-{[4-(4-cyanophenyl)-5-ethoxycarbonyl)pyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carboxylicAcid

tert-Butyl6-[(2-{[4-(4-cyanophenyl)-5-ethoxycarbonyl)pyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carboxylate(prepared in Example 62, 220 mg) was shaken with 100% TFA for 1 hour atroom temperature. The TFA was removed in vacuo. The residue wasdissolved in acetonitrile and water was added. No precipitate formed.Several drops of concentrated ammonium hydroxide were added. Thenglacial acetic acid was added dropwise until white solid formed. Themixture was then filtered to give the title compound as a white solid(180 mg, after drying).

MS: MH⁺=433 C₂₂H₂₀N₆O₄=432 g/mol

NMR (300 MHZ, DMSO-d₆): 8.80 (s, 1H), 8.58 (s, 1H), 7.85 (d, 2H), 7.80(m, 1H), 7.60 (d, 2H), 6.50 (d, 1H), 4.05 (q, 2H), 3.55 (m, 4H), 1.05(t, 3H).

Example 71 Synthesis of Methyl6-[(2-{[4-(4-cyanophenyl)-5-ethoxycarbonyl)pyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carboxylate

Step 1:6-[(2-{[4-(4-cyanophenyl)-5-ethoxycarbonyl)pyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carboxylicacid (prepared in Example 70, 120 mg) was dissolved in thionyl chloride(3 ml) and then warmed at 50° C. for 0.5 hours. The solvent was removedin vacuo to give crude ethyl2-({2-[(5-(chlorocarbonyl)(2-pyridyl))amino]ethyl}amino)-4-(4-cyanophenyl)pyrimidine-5-carboxylate.This compound was dissolved in dry dichloromethane (4 ml).

Step 2: The acid chloride solution prepared in Step 1 (1.0 ml) wastreated with dry methanol (1 ml). After standing approximately 1 hour atroom temperature, the solvent was removed in vacuo to give the titlecompound.

HPLC: 20.90 min (95% purity)

MS: MH⁺=447 C₂₃H₂₂N₆O₄=446 g/mol

Example 72 Synthesis of Ethyl4-(4-cyanophenyl)-2-[(2-{[5-(morpholin-4-ylcarbonyl)(2-pyridyl)]amino}ethyl)amino]pyrimidine-5-carboxylate

The solution of ethyl2-({2-[(5-(chlorocarbonyl)(2-pyridyl))amino]ethyl}amino)-4-(4-cyanophenyl)pyrimidine-5-carboxylatein dichloromethane prepared in Step 1 of Example 71 (1.0 ml) was treatedat room temperature with a solution of morpholine (150 μL) indichloromethane (1 ml). After 1 hour the solvent was removed in vacuo togive the title compound.

HPLC: 19.63 min (96% purity)

MS: MH⁺=502 C₂₆H₂₇N₇O₄=501 g/mol.

Example 73 Synthesis of Ethyl4-(4-cyanophenyl-2-{[2-({5-nitro-6[benzylamino](2-pyridyl)}amino)ethyl]amino}pyrimidine-5-carboxylate

Step 1: 6-Chloro-3-nitro(2-pyridyl))benzylamine was prepared inaccordance with the method described in von Bebenberg, Chemiker-Zeitung,103:387 (1979), which is incorporated herein by reference. This amine(1.8 g) was heated with ethylenediamine (5 ml) in acetonitrile (15 ml)at 100° C. for 3.5 hours. The solvent was removed in vacuo and theresidue was partitioned between dichloromethane and 2.5 M aqueous sodiumhydroxide solution. The aqueous layer was extracted 3× further withdichloromethane. The combined organic layers were washed with asaturated sodium chloride solution, dried and concentrated in vacuo togive a yellow solid.

NMR (300 MHz, CDCl₃): 8.10 (d, 1H), 7.2-7.4 (m, 5H), 5.80 (s, 1H), 4.80(AB q, 2H), 3.42 (m, 2H), 2.85 (m, 2H).

Step 2: The amine from Step 1 (1.31 g) was treated with benzotriazolecarboxamidinium 4-methylbenzenesulfonate (1.52 g) and DIEA (800 μl) inacetonitrile (15 ml) at room temperature overnight. The mixture wasdiluted with ether, then filtered to give the guanidine,amino[2-({5-nitro-6-[benzylamino](2-pyridyl)}amino)ethyl]carboxamidinium4-methylbenzenesulfonate, as a yellow solid.

NMR (300 MHz, DMSO-d₆): 8.02 (d, 1H), 7.72 (d, 2H), 7.30-7.40 (m, 5H),7.10 (d, 2H), 6.00 (d, 1H), 4.78 (AB q, 2H), 3.50 (m, 2H), 3.30 (m, 2H),2.25 (s, 3H).

Step 3: Ethyl 3-(4-cyanophenyl)-3-oxopropanoate (65 mg, 0.3 mmol) washeated with DMFDMA (60 μL) in THF (1 ml) at 70° C. for 3 h. Thissolution was then added to a mixture of the guanidine prepared in Step 2(150 mg, 0.3 mmol), dry ethanol (1 ml) and 1.0 M sodium ethoxide inethanol (0.35 ml) and heated at 80° C. overnight. The solvents wereremoved in vacuo. The residue was partitioned between dichloromethaneand saturated aqueous sodium bicarbonate solution. The organic layer wasconcentrated in vacuo. The residue was dissolved in acetonitrile.Addition of water gave the title compound as a yellow solid.

HPLC: 34.06 min (98% purity)

MS: MH⁺=539 C₂₈H₂₆N₈O₄=538 g/mol

Example 74 Preparation of[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine

1. Preparation of 1-(2,4-dichlorophenyl)-2-imidazol-2-ylethan-1-one.

A solution of 2,4-dichlorobenzoyl chloride (9.75 M) in dichloromethane(75 ml) was added dropwise over 30 minutes to a stirred solution of2-methylimidazole (2) (0.49 M) in dichloromethane (500 ml) andN,N-diisopropylethylamine (Hünig's base) (136 ml). (Ref. Macco, A. A.;Godefroi, E. F.; Drouen, J. J. M., J. Org. Chem. 1975, 40, 252-255.) Thereaction mixture was cooled during the addition using an ice-water bath.The reaction mixture was then heated to reflux for 3.5 hours. A thickreddish black mixture forms. To dilute the thick heterogeneous reaction,additional dichloromethane (100-200 ml) was added as needed to maintainstirring. On cooling dichloromethane (500 ml) was added, and thesolution was transferred to a separatory funnel. The organic layer waswashed with distilled water (3×200 ml). An emulsion formed that breaksapart on sitting for 15 minutes or by filtering. The wet organic layerwas directly concentrated under reduced pressure without drying. Thesolid product was then dried in vacuo for several hours.

To the dry solid (described above) was added a solution (2:1 v/v,500-600 ml) of gla. acetic acid and aq. con. HCl. The mixture was thenstirred at reflux for ca. 75 min. The acetic acid was removed via rotaryevaporator. Distilled water (800 ml) and benzene (400 ml) were added tothe solid residue which was vigorously stirred for 15 min. The solidswere filtered off, and the filtrate was transferred to a separatoryfunnel. After the organic layer was discarded, the aqueous layer waswashed with benzene (4×150 ml). The aqueous layer was transferred to alarge beaker (4 L) and diluted with isopropyl ether (100 ml). Thestirred mixture was basified (pH 7-8) by careful addition of sodiumbicarbonate which leads to the formation of a white solid. After 2 hoursof additional stirring the desired solid was filtered, washed withdistilled water (3×60 ml), isopropyl ether (2×60 ml), and dried in vacuoovernight giving 1-(2,4-dichlorophenyl)-2-imidazol-2-ylethan-1-one in56% yield.

2. Preparation of(2Z)-1-(2,4-dichlorophenyl)-3-(dimethylamino)-2-imidazol-2-ylprop-2-en-1-one.

A mixture of 1-(2,4-dichlorophenyl)-2-imidazol-2-ylethan-1-one (0:39 M)in N,N-dimethylformamide dimethyl acetal (DMFDMA) (150 ml) was stirredfor 2.5 h at 70-75° C. The DMFDMA was then removed under reducedpressure and dried under high vacuum for several hours giving anorange-yellow solid 4 in quantitative yield. The product(2Z)-1-(2,4-dichlorophenyl)-3-(dimethylamino)-2-imidazol-2-ylprop-2-en-1-onewas typically used without further purification.

3. Preparation ofamino{2-[(5-nitro(2-pyridyl))amino]ethyl}carboxamidine, hydrochloride.

A mixture of 2-(2-aminoethylamino)-5-nitropyridine (0.47 M) (purchasedfrom Aldrich, or made by reacting ethylene diamine with2-chloro-5-nitropyridine as per the procedure in example{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amineor example6-[(2-{[4-(2,4-dichlorophenyl)-5-(4-methylimidazol-2-yl)pyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carbonitrile)and 1H -pyrazole-1-carboxamidine hydrochloride (0.47 M) in acetonitrile(500 ml) were stirred overnight (ca. 20 h) at 70-80° C. Upon cooling, ayellow precipitate was collected by filtration. The yellow solid waswashed thoroughly with acetonitrile (3×100 ml), ethyl ether (3×100 ml),and dried overnight in vacuo resulting in 87% yield ofamino{2-[(5-nitro(2-pyridyl))amino]ethyl}carboxamidine, hydrochloride.

4. Preparation of[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine.

A solution of sodium ethoxide (0.59 M) dissolved in abs. ethanol (100ml) was added to a stirred mixture of(2Z)-1-(2,4-dichlorophenyl)-3-(dimethylamino)-2-imidazol-2-ylprop-2-en-1-one(0.23 M) and amino{2-[(5-nitro(2-pyridyl))amino]ethyl}carboxamidine,hydrochloride (0.23 M) in abs. ethanol (260 ml). The reaction wasstirred at room temperature for 15 min. and then at 75-80° C. for 2.5hours. On cooling a yellow precipitate was collected by filtration. Thefiltrate was stored for possible further isolation and purification ofthe product. The solid product was washed with abs. ethanol (3×50 ml),distilled water (3×50 ml), and ethyl ether (3×50 ml). The yellow solidwas dried overnight in vacuo giving 52.7% yield of final product[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC: 20.9 min (>95% purity)

MS: M+H=471 (C₂₀H₁₆C₁₂N₈O₂+H=471)

Example 75 Preparation of{2-[(4-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amine

{2-[(4-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]aminewas prepared using the general method for[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine(see Example 74) with the exceptions noted below.

1. Preparation of 2-(2-aminoethyl)amino-6-amino-5-nitropyridine.

A mixture of 2-amino-6-chloro-3-nitropyridine (0.52 M) in acetonitrile(70 ml), and ethylene diamine (40 ml) were stirred overnight (ca. 20 h)at 75-80° C. under argon. The ethylene diamine was removed under reducedpressure. The residual solution was basified with 1M sodium hydroxidesolution (50 ml). The aqueous solution was saturated with sodiumchloride and extracted with a solution of 95% ethyl acetate and 5%methanol (3×150 ml) and with a solution of 95% acetonitrile and 5%methanol (3×150 ml). The organic extracts were combined and extractedwith a saturated sodium chloride solution (2×75 ml). The organic layerwas dried with sodium sulfate, filtered, and concentrated under reducedpressure. The crude yellow solid was triturated with ether (2×25 ml) anddried overnight in vacuo resulting in2-(2-aminoethyl)amino-6-amino-5-nitropyridine in 99% yield.

2. Preparation ofamino{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}carboxamidine,hydrochloride.

A mixture of 2-(2-aminoethyl)amino-6-amino-5-nitropyridine (0.44 M), 1H-pyrazole-1-carboxamidine hydrochloride (0.44 M) in acetonitrile (75 ml)were stirred overnight (ca. 24 h) at 75-80° C. Upon cooling, a yellowprecipitate was collected by filtration. The yellow solid was washedthoroughly with acetonitrile (3×50 ml), ethyl ether (3×50 ml), and driedovernight in vacuo givingamino{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}carboxamidine as theHCl salt in 82% yield.

3. Preparation of{2-[(4-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amine.

A solution of sodium ethoxide (0.5 M) dissolved in abs. ethanol (8 ml)was added to a stirred mixture of(2Z)-1-(2,4-dichlorophenyl)-3-(dimethylamino)-2-imidazol-2-ylprop-2-en-1-one(0.57 M),amino{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}carboxamidine,hydrochloride (0.58 M), and abs. ethanol (7 ml). The reaction was thenheated to 75-80° C. for 2.5 hours. On cooling a yellow precipitate wascollected by filtration. The filtrate was stored for possible furtherisolation and purification of final product. The solid product waswashed with abs. ethanol (3×10 ml), distilled water (3×10 ml), and ethylether (3×10 ml). The yellow solid was dried overnight in vacuo giving{2-[(4-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]aminein70% yield.

HPLC: 18.7 min (>95% purity)

MS: M+H=486.2 (C₂₀H₁₇Cl₂N₉O₂+H=486)

Example 76 Preparation of6-[(2-{[4-(4-cyanophenyl)-5-imidazolylpyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carbonitrile

1. Preparation of 4-(2-imidazolylacetyl)benzenecarbonitrile.

A stirred solution of 4-(2-chloroacetyl)benzenecarbonitrile (0.5 M) andimidazole (1.5 M) in CH₃CN (200 ml) were heated for 14 hours at 60° C.The product was stripped of solvent under reduced pressure. The residuewas diluted with dichloromethane (250 ml) and water (100 ml), and themixture was stirred for 30 min. After filtering off a solid impurity,the aqueous layer was removed and discarded. The organic layer waswashed sequentially with water (60 ml), sat. aq. NaHCO₃ (60 ml), water(60 ml), brine (60 ml), dried with Na₂SO₄, filtered, and concentratedunder reduced pressure. The dark colored oil was dried overnight invacuo giving 4-(2-imidazolylacetyl)benzenecarbonitrile in 90% yield.

2. Preparation of4-[(2E)-3-(dimethylamino)-2-imidazolylprop-2-enoyl]benzene-carbonitrile.

A mixture of 4-(2-imidazolylacetyl)benzenecarbonitrile (0.30 M) andN,N-dimethylformamide dimethyl acetal (DMFDMA) (80 ml) was stirred for12 h at 75° C. The DMFDMA was then removed under reduced pressure anddried under high vacuum for several hours giving an orange-yellow solid4-[(2E)-3-(dimethylamino)-2-imidazolylprop-2-enoyl]benzenecarbonitrilein quantitative yield. The enaminone product was typically used withoutfurther purification.

3. Preparation of amino{2-[(5-cyano(2-pyridyl))amino]ethyl}carboxamidine, hydrochloride).

The preparation of the materialamino{2-[(5-cyano(2-pyridyl))amino]ethyl}carboxamidine, hydrochloridecan be found in the procedures for the preparation of6-[(2-{[4-(2,4-dichlorophenyl)-5-(4-methylimidazol-2-yl)pyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carbonitrile.

4. Preparation of6-[(2-{[4-(4-cyanophenyl)-5-imidazolylpyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carbonitrile.

A solution of sodium ethoxide (0.66 M) dissolved in abs. ethanol (8 ml)was added to a stirred mixture of4-[(2E)-3-(dimethylamino)-2-imidazolylprop-2-enoyl]benzenecarbonitrile(0.33 M), amino{2-[(5-cyano(2-pyridyl))amino]ethyl}carboxamidine,hydrochloride (0.33 M), and abs. ethanol (15 ml). The reaction was thenheated to 75-80° C. for 2.5 hours. On cooling the reaction was dilutedwith ethyl acetate (400 ml) washed with sat. aq. NaHCO₃ (100 ml),distilled water (2×100 ml), brine (100 ml), dried with Na₂SO₄, filtered,and concentrated. The crude product was purified by flash chromatographyover silica gel. The column was run starting with 1:1 ethyl acetate tohexane, then ethyl acetate which was used until all of the fast movingimpurities had been removed. The product was eluted with 1.5% methanolin ethyl acetate. The column is monitored by TLC using 5% methanol inethyl acetate as the solvent system. The proper fractions werecondensed. The off-white solid was dried overnight in vacuo giving6-[(2-{[4-(4-cyanophenyl)-5-imidazolylpyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carbonitrilein 40% yield.

HPLC: 18.9 min (>95% purity)

MS: M+H=473.1 (C₂₂H₁₇N₉+H=473)

Example 77 Preparation of(tert-butoxy)-N-(2-{[4-(4-cyanophenyl)-5-imidazolylpyrimidin-2-yl]amino}ethyl)carboxamide

1. Preparation of N-(2-aminoethyl)(tert-butoxy)carboxamide.

A solution of tert-butyl[(tert-butyl)oxycarbonyloxy]formate (Boc₂O)(181g, 830 mmol) in dichloromethane (1 L) was added slowly to amechanically stirred solution of ethylene diamine (250 g, 4.16 mol) indichloromethane (2.5 L) at room temperature. After 24 hours, thereaction solution was washed with water (3×500 ml), Brine (500 ml),dried with Na₂SO₄, filtered, and concentrated under reduced pressure.The pure product N-(2-aminoethyl)(tert-butoxy)carboxamide was obtainedin 50% yield.

2. Preparation of N-[2-(amidinoamino)ethyl](tert-butoxy)carboxamide,hydrochloride.

Portions of solid 1H-pyrazole-1-carboxamidine hydrochloride (91.10 g,624 mmol)) are added to a stirred solution ofN-(2-aminoethyl)(tert-butoxy)carboxamide (100 g, 624 imol) in CH₃CN (1L) at 80° C. After 24 hours, the reaction was stripped of solvent underreduced pressure. The residue was triturated with ether (3 x 100 ml),and dried in vacuo. The guanidineN-[2-(amidinoamino)ethyl](tert-butoxy)carboxamide, hydrochloride wasobtained in over 100% yield containing a small amount of pyrazole. Theguanidine was used without further purification.

3. Preparation of(tert-butoxy)-N-(2-{[4-(4-cyanophenyl)-5-imidazolylpyrimidin-2-yl]amino}ethyl)carboxamide.

4-[(2E)-3-(dimethylamino)-2-imidazolylprop-2-enoyl]benzenecarbonitrile(8.0g, 30.0 mmol) in NMP (5 ml) was added to a stirred mixture ofN-[2-(amidinoamino)ethyl](tert-butoxy)carboxamide, hydrochloride (13.8g, 45 =mol)) and Cs₂CO₃ (11.72, 36.0 mmol) in NMP (15 ml). The reactionwas heated to 100° C. for 48 hours. The reaction was followed by HPLC.Upon completion, the reaction was partitioned with water (50 ml) anddichloromethane (250 ml). The organic layer was separated and washedwith water (2×50 ml), brine (50 ml), dried Na₂SO₄, filtered, andconcentrated under reduced pressure. The product was purified by flashchromatography eluting with 10% methanol in dichloromethane. Afterstripping off the solvents and drying in vacuo, 10.08 g of(tert-butoxy)-N-(2-{[4-(4-cyanophenyl)-5-imidazolylpyrimidin-2-yl]amino}ethyl)carboxamidewas obtained as a dark red glass in 83% yield.

Example 78 Preparation of4-{5-imidazolyl-2-[(2-{[5-(trifluoromethyl)(2-pyridyl)]amino}ethyl)amino]pyrimidin-4-yl}benzenecarbonitrile

4-{5-imidazolyl-2-[(2-{[5-(trifluoromethyl)(2-pyridyl)]amino}ethyl)amino]pyrimidin-4-yl}benzenecarbonitrilewas prepared using the general method for6-[(2-{[4-(4-cyanophenyl)-5-imidazolylpyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carbonitrile(60406) (see Example 76) with the exceptions noted below.

1. Preparation of N-(2-aminoethyl)(tert-butoxy)carboxamide.

A solution of tert-butyl [(tert-butyl)oxycarbonyloxy]formate (Boc₂O)(0.83 M) in dichloromethane (1 L) was added slowly (3 h) to amechanically stirred solution of ethylene diamine (1.66 M) indichloromethane (2.5 L) at room temperature. After 24 hours, thereaction solution was washed with water (3×500 ml), brine (500 ml),dried with Na₂SO₄, filtered, and concentrated under reduced pressure.The pure N-(2-aminoethyl)(tert-butoxy)carboxamide was obtained in 50%yield.

2. Preparation of N-[2-(amidinoamino)ethyl](tert-butoxy)carboxamide,hydrochloride.

Portions of solid 1H-pyrazole-1-carboxamnidine hydrochloride (91.10 g,624 mmol)) are added to a stirred solution ofN-(2-aminoethyl)(tert-butoxy)carboxamide (0.62 M) in CH₃CN (1 L) at 80°C. After 24 hours, the reaction was stripped of solvent under reducedpressure. The residue was triturated with ether (3×100 ml), and dried invacuo. The N-[2-(amidinoamino)ethyl](tert-butoxy)carboxamide,hydrochloride was obtained in over 100% yield containing a small amountof pyrazole. The N-[2-(amidinoamino)ethyl](tert-butoxy)carboxamide,hydrochloride was used without further purification.

3. Preparation of(tert-butoxy)-N-(2-{[4-(4-cyanophenyl)-5-imidazolylpyrimidin-2-yl]amino}ethyl)carboxatnide.4-[(2E)-3-(dimethylamino)-2-imidazolylprop-2-enoyl]benzenecarbonitrile(6 M) in NMP (5 ml) was added to a stirred mixture ofN-[2-(amidinoamino)ethyl](tert-butoxy)carboxarnide, hydrochloride (3 M)and Cs₂CO₃ (2.4) in NMP (15 ml). The reaction was heated to 100° C. for48 hours. The reaction was followed by HPLC. Upon completion, thereaction was partitioned with water (50 ml) and dichloromethane (250ml). The organic layer was separated and washed with water (2×50 ml),brine (50 ml), dried Na₂SO₄, filtered, and concentrated under reducedpressure. The product was purified by flash chromatography eluting with10% methanol in dichloromethane. After stripping off the solvents anddrying in vacuo,(tert-butoxy)-N-(2-{[4-(4-cyanophenyl)-5-imidazolylpyrimidin-2-yl]amino}ethyl)carboxamidewas obtained as a dark red glass in 83% yield.4. Preparation of4-{2-[(2-aminoethyl)amino]-5-imidazolylpyrimidin-4-yl}benzenecarbonitrile.

Aqueous 3M HCl (15-30 ml) was added to a stirred solution of(tert-butoxy)-N-(2-{[4-(4-cyanophenyl)-5-imidazolylpyrimidin-2-yl]amino}ethyl)carboxamide(0.15 M) in CH₃CN (50 ml) at room temperature until the reaction becameslightly turbid. After 16 hours, the reaction was partitioned betweendichloromethane (200 ml) and 1M HCl (200 ml). The layers were separated,and the aqueous layer was extracted with dichloromethane (3×200 ml). Theaqueous layer was basified carefully with solid NaHCO₃ to pH 7-8. Asolid forms which can be filtered and dissolved in CH₃CN (100 ml). Theorganic solution is washed with sat. aq. NaHCO₃ (50 ml), brine (50 ml),dried with Na₂SO₄, filtered, and concentrated. The crude product waspurified by flash chromatography over silica gel. The column was elutedfirst with a 1:1 mixture of dichloromethane/methanol followed by amixture of 5% TEA/10% water/85% methanol to elute the product. Theproper fractions were condensed. The dark yellow glass was driedovernight in vacuo giving4-{2-[(2-aminoethyl)amino]-5-imidazolylpyrimidin-4-yl}benzenecarbonitrilein 89% yield.

5. Preparation of4-{5-imidazolyl-2-[(2-{[5-(trifluoromethyl)(2-pyridyl)]amino}ethyl)amino]pyrimidin-4-yl}benzenecarbonitrile.

A mixture of4-{2-[(2-aminoethyl)amino]-5-imidazolylpyrimidin-4-yl}benzene-carbonitrile(30 mg, 0.098 mmol), 2-chloro-5-(trifluoromethyl)pyridine (18 mg, 0.098mmol), and Hünig's base (70 uL, 0.4 mmol) in DMA (500 uL) were heated to80° C. After stirring for 12 hours, the reaction was diluted with ethylacetate (10 ml) and extracted with sat. aq. NaHCO₃ (2×5 ml), water (3×5ml), brine (5 ml), dried with Na₂SO₄, filtered, and concentrated. Theproduct was purified by preparative HPLC using a reverse phase columnand a water/acetonitrile gradient. The product4-{5-imidazolyl-2-[(2-{[5-(trifluoromethyl)(2-pyridyl)]amino}ethyl)amino]pyrimidin-4-yl}benzenecarbonitrilewasobtained as a off-white solid in 5% yield.

HPLC: 16.3 min (>95% purity)

MS: M+H=451.2 (C₂₂H₁₇F₃N₈+H=451)

Example 79 Preparation of4-{5-imidazolyl-2-[(2-{[(4-nitrophenyl)sulfonyl]amino}ethyl)amino]pyrimidin-4-yl}benzenecarbonitrile

A mixture of4-{2-[(2-aminoethyl)amino]-5-imidazolylpyrimidin-4-yl}benzene-carbonitrile(30 mg, 0.098 mmol), chloro(4-nitrophenyl)sulfone (22 mg, 0.1 nmuol),and Hünig's base (70 uL, 0.4 mmol) in DMA (500 uL) were heated to 80° C.After stirring for 12 hours, the reaction was diluted with ethyl acetate(10 ml) and extracted with sat. aq. NaHCO₃ (2×5 ml), water (3×5 ml),brine (5 ml), dried with Na₂SO₄, filtered, and concentrated. Theresulting product, 4-{5-imidazolyl-2-[(2-{[(4-nitrophenyl)sulfonyl]amino}ethyl)amino]pyrimidin-4-yl}benzenecarbonitrile weighing27 mg (56% yield) was approximately 90% pure by LCMS and HPLC.

Example 80 Preparation ofN-(2-{[4-(4-cyanophenyl)-5-imidazolylpyrimidin-2-yl]amino}ethyl)(3-nitrophenyl)carboxamide

A mixture of4-{2-[(2-aminoethyl)amino]-5-imidazolylpyrimidin-4-yl}benzene-carbonitrile(30 mg, 0.098 mmol), 3-nitrobenzoic acid (17 mg, 0.1 mmol),1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) (19mg, 0.1 mmol), 1-bydroxybenzotriazole hydrate (HOBT) (14 mg, 0.1 mmol),and 4-dimethylaminopyridine (DMAP) (12 mg, 0.1 mmol) in DMA (500 uL)were stirred at room temperature. After 12 hours, the reaction wasdiluted with ethyl acetate (10 ml) and extracted with sat. aq. NaHCO₃(2×5 ml), water (3×5 ml), brine (5 ml), dried with Na₂SO₄, filtered, andconcentrated. The resulting product,N-(2-{[4-(4-cyanophenyl)-5-imidazolylpyrimidin-2-yl]amino}ethyl)(3-nitrophenyl)carboxamideweighing 32 mg (70% yield) was approximately 95% pure by LCMS and HPLC.

Example 81 Preparation of[5-amino-4-(2,4-dichlorophenyl)pyrimidin-2-yl]{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amine

1. Preparation of2-[2-(2,4-dichlorophenyl)-2-oxoethyl]isoindoline-1,3-dione

1 mmol of 2,4-dichlorophenacyl chloride in DMF was added drop wise to 2mmol of phthalimide and 2 mmol of Cs₂CO₃ in DMF at room temperature forfourteen hours and then purified by trituration with diethyl ether toobtain 2-[2-(2,4-dichlorophenyl)-2-oxoethyl]isoindoline-1,3-dione.

2. Preparation of2-{2-(2,4-dichlorophenyl)-1-[(dimethylamino)methylene]-2-oxoethyl}isoindoline-1,3-dione

1 mmol of 2-[2-(2,4-dichlorophenyl)-2-oxoethyl]isoindoline-1,3-dione washeated to 80° C. in neat N,N-dimethylformamidedimethyl acetal for sixhours. The reaction mixture was concentrated in vacuo and purified bytrituration with diethyl ether to obtain2-{2-(2,4-dichlorophenyl)-1-[(dimethylamino)methylene]-2-oxoethyl}isoindoline1,3-dione.

3. Preparation of2-{N-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]carbamoyl}benzoicacid

1 mmol of2-{2-(2,4-dichlorophenyl)-1-[(dimethylamino)methylene]-2-oxoethyl}isoindoline-1,3-dione,1 mmol of amino{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}carboxamidine, and 3 mmol of Cs₂CO₃ were dissolved in DMFand heated to 90° C. for fourteen hours. The reaction mixture wasconcentrated in vacuo and diluted with water and ethyl acetate. Thesolution was extracted three times with ethyl acetate and dried oversodium sulfate to obtain2-{N-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]carbamoyl}benzoicacid.

4. Preparation of2-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]isoindoline-1,3-dione

1 mmol of2-{N-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]carbamoyl}benzoicacid was heated to 120° C. in acetic acid for four hours and thenconcentrated in vacuo to obtain2-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]isoindoline-1,3-dione.

Preparation of[5-amino-4-(2,4-dichlorophenyl)pyrimidin-2-yl]{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amine

1 mmol of2-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]isoindoline-1,3-dioneand 20 mnol of hydrazine were stirred in ethanol at 75° C. for two hoursand the resulting product,[5-amino-4-(2,4-dichlorophenyl)pyrimidin-2-yl]{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}aminewas purified by column chromatography eluting with 5-10%methanol/methylene chloride.

HPLC: 5.704min. (100% purity)

MS:MH⁺=435.1

Example 82 Preparation of{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-morpholin-4-ylpyrimidin-2-yl]amine

1. Preparation of 1-(2,4-dichlorophenyl)-2-morpholin-4-ylethan-1-one

1 mmol of 2,4-dichlorophenacyl chloride in DMF was added drop wise to 10mnmol morpholine in DMF at room temperature for fourteen hours. Thereaction mixture was concentrated in vacuo and diluted with water andethyl acetate. The solution was extracted three times with ethylacetate, dried over sodium sulfate, and purified by columnchromatography eluting with 50% ethyl acetate and 50% hexane to obtain1-(2,4-dichlorophenyl)-2-morpholin-4-ylethan-1-one.

2. Preparation of(2E)-1-(2,4-dichlorophenyl)-3-(dimethylamnino)-2-morpholin-4-ylprop-2-en-1-one

1 mmol of 1-(2,4-dichlorophenyl)-2-morpholin-4-ylethan-1-one was heatedto 80° C. in N,N-dimethylformamidedimethyl acetal for six hours. Thereaction mixture was concentrated in vacuo and purified by triturationwith diethyl ether to obtain(2E)-1-(2,4-dichlorophenyl)-3-(dimethylamino)-2-morpholin-4-ylprop-2-en-1-one.

3. Preparation of{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-morpholin-4-ylpyrimidin-2-yl]amine

1 mmol of(2E)-1-(2,4-dichlorophenyl)-3-(dimethylamino)-2-morpholin-4-ylprop-2-en-1-one,1 mmol ofamino{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}carboxamidine, and 3mmol of Cs₂CO₃ were dissolved in DMF and heated to 90° C. for fourteenhours. The reaction mixture was concentrated in vacuo and diluted withwater and ethyl acetate. The solution was extracted three times withethyl acetate and purified by column chromatography eluting with 5-10%methanol/methylene chloride toobtain{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-morpholin-4-ylpyrimidin-2-yl]amine.

HPLC: 9.367min. (100% purity)

MS: MH⁺=505

Example 83 Preparation of1-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]pyrrolidine-2,5-dione

1. Preparation of2-[2-(2,4-dichlorophenyl)-2-oxoethyl]isoindoline-1,3-dione

1 mmol of 2,4-dichlorophenacyl chloride in DMF was added drop wise to 2mmol of phthalimide and 2 mmol of Cs₂CO₃ in DMF at room temperature forfourteen hours and then purified by trituration with diethyl ether toobtain 2-[2-(2,4-dichlorophenyl)-2-oxoethyl]isoindoline-1,3-dione.

2. Preparation of2-{2-(2,4-dichlorophenyl)-1-[(dimethylamino)methylene]-2-oxoethyl}isoindoline-1,3-dione

1 mmol of 2-[2-(2,4-dichlorophenyl)-2-oxoethyl]isoindoline-1,3-dione washeated to 80° C. in neat N,N-dimethylformamidedimethyl acetal for sixhours. The reaction mixture is concentrated in vacuo and purified bytrituration with diethyl ether to obtain2-{2-(2,4-dichlorophenyl)-1-[(dimethylamino)methylene]-2-oxoethyl}isoindoline-1,3-dione.

3. Preparation of2-{N-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]carbamoyl}benzoicacid

1 mmol of2-{2-(2,4-dichlorophenyl)-1-[(dimethylamino)methylene]-2-oxoethyl}isoindoline-1,3-dione,1 mmol ofamino{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}carboxamidine, and 3mmol of Cs₂CO₃ were dissolved in DMF and heated to 90° C. for fourteenhours. The reaction mixture was concentrated in vacuo and diluted withwater and ethyl acetate. The solution was extracted three times withethyl acetate and dried over sodium sulfate to obtain2-{N-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]carbamoyl}benzoicacid.

4. Preparation of2-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]isoindoline-1,3-dione

1 mmol of2-{N-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]carbamoyl}benzoicacid is heated to 120° C. in acetic acid for four hours and thenconcentrated in vacuo to obtain2-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]isoindoline-1,3-dione.

5. Preparation of[5-amino-4-(2,4-dichlorophenyl)pyrimidin-2-yl]{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amine

1 mmol of2-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]isoindoline-1,3-dioneand 20 mmol of hydrazine were stirred in ethanol at 75° C. for two hoursand then purified by column chromatography eluting with 5-10%methanol/methylene chloride to obtain[5-amino-4-(2,4-dichlorophenyl)pyrimidin-2-yl]{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amine.

6. Preparation of1-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]pyrrolidine-2,5-dione

1 mmol of[5-amino-4-(2,4-dichlorophenyl)pyrimidin-2-yl]{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amine,and 2 mmol of succinic acid, 4 mmol of HBTU, and 5 mmol ofN,N-diisopropylethylamine were added to solution and stirred for sixhours at room temperature. The reaction mixture was concentrated invacuo and diluted with water and ethyl acetate. The solution wasextracted three times with ethyl acetate, dried over sodium sulfate, andpurified by column chromatography 5-10% methanol/methylene chloride toobtain1-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]pyrrolidine-2,5-dione.

HPLC: 8.917min. (100% purity)

MS: MH+=517.1

Example 84 Preparation of{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-piperazinylpyrimidin-2-yl]amine

1. Preparation of tert-butyl4-[2-(2,4-dichlorophenyl)-2-oxoethyl]piperazinecarboxylate

1 mmol of 2,4-dichlorophenacyl chloride in DMF was added drop wise to1.2 mmol of tert-butyl piperazinecarboxylateand 1.2 mmol Cs₂CO₃ in DMFat room temperature for fourteen hours. The reaction mixture wasconcentrated in vacuo and diluted with water and ethyl acetate. Thesolution was extracted three times with ethyl acetate, dried over sodiumsulfate, and purified by column chromatography eluting with 50% ethylacetate and 50% hexane to obtain tert-butyl4-[2-(2,4-dichlorophenyl)-2-oxoethyl]piperazinecarboxylate.

2. Preparation oftert-butyl4-{2-(2,4-dichlorophenyl)-1-[(dimethylamino)methylene]-2-oxoethyl}piperazinecarboxylate

1 mmol of tert-butyl4-[2-(2,4-dichlorophenyl)-2-oxoethyl]piperazinecarboxylate was heated to80° C. in neat N,N-dimethylformamidedimethyl acetal for six hours. Thereaction mixture was concentrated in vacuo and purified by triturationwith diethyl ether to obtain tert-butyl4-{2-(2,4-dichlorophenyl)-1-[(dimethylamino)methylene]-2-oxoethyl}piperazinecarboxylate.

3. Preparation of tert-butyl4-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]piperazinecarboxylate

1 mmol of tert-butyl4-{2-(2,4-dichlorophenyl)-1-[(dimethylamino)methylene]-2-oxoethyl}piperazinecarboxylate,1 mmol of amino{2-[(5-nitro(2-pyridyl))amino]ethyl}carboxamidine, and 3mmol of Cs₂CO₃ were dissolved in DMF and heated to 90° C. for fourteenhours. The reaction mixture was concentrated in vacuo and diluted withwater and ethyl acetate. The solution was extracted three times withethyl acetate, dried over sodium sulfate, and purified by columnchromatography eluting with 5-10% methanolmethylene chloride to obtaintert-butyl4-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]piperazinecarboxylate.

4. Preparation of{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-piperazinylpyrimidin-2-yl]amine

1 mmol of tert-butyl4-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]piperazinecarboxylatewas heated to 60° C. 3M HCl in MeOH for one hour and concentrated invacuo toobtain{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-piperazinylpyrimidin-2-yl]amine.

HPLC: 5.27min. (100% purity)

MS: MH⁺=504.2

Example 85 Preparation of{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(4-ethylphenyl)-5-imidazolylpyrimidin-2-yl]amine

1. Preparationof2-bromo-1-(4-ethylphenyl)ethan-1-one

20 mmol of 1-(4-ethylphenyl)ethan-1-one, 1 ml conc. HCl were mixed in 20ml diethyl ether at 0° C. under nitrogen. To this solution a solution of20 mmol Br₂ in 20 ml chloroform was added drop wise and left for fourhours and then concentrated in vacuo to obtain2-bromo-1-(4-ethylphenyl)ethan-1-one.

2. Preparation of 1-(4-ethylphenyl)-2-imidazolylethan-1-one

1 mmol of 2-bromo-1-(4-ethylphenyl)ethan-1-one in acetonitrile was addeddrop wise to 5.5 mmol imidazoline in acetonitrile at room temperaturefor fourteen hours. The reaction mixture was concentrated in vacuo anddiluted with water and ethyl acetate. The solution was extracted threetimes with ethyl acetate and dried over sodium sulfate to obtain1-(4-ethylphenyl)-2-imidazolylethan-1-one.

3. Preparation of(2E)-3-(dimethylamino)-1-(4-ethylphenyl)-2-imidazolylprop-2-en-1-one

1 mmol of 1-(4-ethylphenyl)-2-imidazolylethan-1-one was heated to 80° C.in N,N-dimethylformamidedimethyl acetal for six hours and concentratedin vacuo to obtain(2E)-3-(dimethylamino)-1-(4-ethylphenyl)-2-imidazolylprop-2-en-1-one.

4. Preparationof{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(4-ethylphenyl)-5-imidazolylpyrimidin-2-yl]amine

1 mmol of(2E)-3-(dimethylamino)-1-(4-ethylphenyl)-2-imidazolylprop-2-en-1-one, 1mmol of amino{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}carboxamidine,and 3 mmol of CS₂CO₃ was dissolved in DMF and heated to 90° C. forfourteen hours. The reaction mixture was concentrated in vacuo anddiluted with water and ethyl acetate. The solution was extracted threetimes with ethyl acetate, dried over sodium sulfate, and purified bycolumn chromatography eluting with 5-10% methanol/methylene chloride toobtain{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(4-ethylphenyl)-5-imidazolyl-pyrimidin-2-yl]amine.

HPLC: 7.733min. (100% purity)

MS: MH⁺=446.2

Example 86 Preparation of{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-(4-pyridyl)pyrimidin-2-yl]amine

1-(2,4-dichlorophenyl)-2-(4-pyridyl)ethan-1-one was synthesized as inSuzuki et al., “Facile dibenzoylation of picoline,” J HeterocycL. Chem.22(6):1487-9 (1985). 1 mmol of1-(2,4-dichlorophenyl)-2-(4-pyridyl)ethan-1-one was heated to 80° C. inneat DMF-DMA for six hours. The reaction mixture was concentrated invacuo and the residue was purified by trituration with diethyl ether.using amino{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}carboxamidinehydrochloride (160mg, 0.58mmol, 1 eq) and the enaminone(2E)-1-(2,4-dichlorophenyl)-3-(dimethylamino)-2-(4-pyridyl)prop-2-en-1-one(202mg, 0.58mmol) and Cs₂CO₃(246mg, 1.3eq) in 5ml DMF at 95° C. for 6h.It was worked up by concentration in vacuo and extraction into ethylacetate to yield a crude product that was purified by columnchromatography, eluting with 10% methanol in methylene chloride to yield{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-(4-pyridyl)pyrimidin-2-yl]amineas a yellow powder after lyophilization.

HPLC: 6.32 min(100%)

MS:MH⁺=497

Example 87 Preparation of{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}{4-(2,4-dichlorophenyl)-5-[5-(trifluoromethyl)(1,2,3,4-tetraazolyl)]pyrimidin-2yl}amine

Trifluoromethyl tetrazole was made according to the procedure publishedin Tetrahedron Letters 38(7):1257-1260. A mixture of tetrachlorosilane(5mmol) and sodium azide (15mmol) in dry acetonitrile (10 ml) andtrifluoromethyl acetamide (5mmol) was refluxed and stirred with theexclusion of moisture. The reaction mixture was poured into ice coldsodium carbonate solution and extracted with chloroform (3×20 ml). Thesolvent was distilled off under reduced pressure to yieldtrifluoromethyl tetrazole (MS:MH−=136.7) which was used without furtherpurification. Trifluromethyl tetrazole (1 mmol), Cs₂CO₃ (1.3 mmol) and1-(2,4-dichlorophenyl)-2-chloroethan-1-one (1 mmol) were refluxed inDMF(2ml) overnight. The reaction mixture was cooled and concentrated invacuo, and then extracted into ethyl acetate and dried over sodiumsulfate. The extract was further purified by column chromatography onsilica gel to yield1-(2,4-dichlorophenyl)-2-[5-(trifluoromethyl)(1,2,3,4-tetraazolyl)]ethan-1-one.1 mmol of1-(2,4-dichlorophenyl)-2-[5-(trifluoromethyl)(1,2,3,4-tetraazolyl)]ethan-1-onewas heated to 80° C. in neat DMF-DMA for six hours. The reaction mixturewas concentrated in vacuo and purified by trituration with diethylether. 1 mmol of the above enaminone, 1 mmol ofamino{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}carboxamidinehydrochloride and 3 mmol of Cs₂CO₃ were dissolved in DMF and heated to90° C. for fourteen hours. The reaction mixture was concentrated invacuo and extracted into ethyl acetate and the residue after solventevaporation was purified by column chromatography eluting with 5-10%methanol in methylene chloride toyield{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}{4-(2,4-dichlorophenyl)-5-[5-(trifluoromethyl)(1,2,3,4-tetraazolyl)]pyrimidin-2-yl}amineas a yellow powder after lyophilization.

MS:MH⁺=556.0,

HPLC: 10.77min (98.3%)

Example 88 Preparation of{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-(4-methylpiperazinyl)pyrimidin-2-yl]amine

1-(2,4-dichlorophenyl)-2-chloroethan-1-one(1 mmol) and methylpiperazine(4 mmol) in 8ml DMF was stirred for 8h at room temperature. The reactionmixture was concentrated in vacuo and extracted into ethyl acetate andthe organic layer was washed with water and dried with sodium sulfate.The residue after concentration in vacuo was purified by columnchromatography eluting with 10% methanol in methylene chloride to yield1-(2,4-dichlorophenyl)-2-(4-methylpiperazinyl)ethan-1-one. The ethanonewas taken up in DMF-DMA and heated to 80° C. for 6h. The reactionmixture was cooled and concentrated in vacuo and the residue containingthe enaminone was purified by column chromatography. 1 mmol of theenaminone obtained above, 1 mmol ofamino{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}carboxamidinehydrochloride and 3 mmol of Cs₂CO₃ was suspended in DMF and heated to90° C. for fourteen hours. The reaction was cooled and concentrated invacuo. The residue was partitioned between water and ethyl acetate andthe layers separated. The organic layers were dried with sodium sulfateand after solvent removal were chromatographed on silica gel elutingwith 5-10% methanol in methylene chloride toobtain{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-(4-methylpiperazinyl)pyrimidin-2-yl]amineas a yellow powder after lyophilization.

LC RT 5.193 min (95.3%)

MS:MH⁺=518.2

Example 89 Preparation of[4-(2,4-dichlorophenyl)-5-(4-methylpiperazinyl)pyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine

1-(2,4-dichlorophenyl)-2-chloroethan-1-one(1mmol) and methylpiperazine(4mmol) in 8ml DMF was stirred for 8h at room temperature. The reactionmixture was then concentrated in vacuo and extracted into ethyl acetateand the organic layer was washed with water and dried with sodiumsulfate. The residue after concentration in vacuo was purified by columnchromatography eluting with 10% methanol in methylene chloride to yield1-(2,4-dichlorophenyl)-2-(4-methylpiperazinyl)ethan-1-one.1-(2,4-dichlorophenyl)-2-(4-methylpiperazinyl)ethan-1-one was heated to80° C. in neat DMF-DMA for six hours. The reaction mixture wasconcentrated in vacuo and the residue obtained was purified bytrituration with diethyl ether to yield the enaminone. 1 mmol of theenaminone obtained above, 1mmol ofamino{2-[(5-nitro(2-pyridyl))amino]ethyl}carboxamidine hydrochloride and1.3 mmol of Cs₂CO₃ were dissolved in DMF and heated to 90° C. forfourteen hours. The reaction mixture was cooled to room temperature andconcentrated in vacuo. Water was added and then extracted 3× with ethylacetate. The combined organic layers were dried with sodium sulfate. Theresidue after solvent evaporation was purified by column chromatographyeluting with 5-10% methanol in methylene chloride to yield[4-(2,4-dichlorophenyl)-5-(4-methylpiperazinyl)pyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amineas a yellow powder after lyophilization.

HPLC 5.933min (>95%)

MS:MH⁺=503.1

Example 90 Preparation of4-[6-imidazolyl-2-({2-[(5-nitro(2-pyridyl))aminolethyl}amino)pyrimidin-4-yl]benzenecarbonitrile

According to the procedure of Wierenga (Heterocycles, 23:1687 (1985)),9.45 g (0.034 mol) of S-methyl isourea nitrate, 13.57g (0.062 mol) ofethyl 3-(4-cyanophenyl)-3-oxopropanoate and 8.0 g (0.142 mol) ofpotassium hydroxide were suspended in 40 ml water and the heterogeneousmixture stirred for 18 h. The mixture was then filtered, and the solidwashed with copious amounts of water. The solid was dried, andrecrystalized from 5% dimethylformamide/ethanol to afford 2.47 g (34%)of 4-(2-amino-4-oxo-1,3-oxazin-6-yl)benzenecarbonitrile as a colorlesssolid.

A solution of 44 ml DMF, 3.81 g (17.89 mmol)4-(2-amino-4-oxo-1,3-oxazin-6-yl)benzenecarbonitrile and 8.57 g (53.63mmol) of N-(2-aminoethyl) (tert-butoxy)carboxamide were heated to 50° C.for 3 h. The reaction was then concentrated in vacuo (0.5 mm Hg) and theresulting yellow solid triturated with cold ethanol. The resultingsuspension was filtered and the solid washed with copious amounts ofethanol. This gave 4.51 g (71%) of(tert-butoxy)-N-(2-{[6-(4-cyanophenyl)-4-hydroxypyrimidin-2-yl]amino}ethyl)carboxamideas a white solid.

To a solution of 4.01 g (11.2 mmol) of(tert-butoxy)-N-(2-{[6-(4-cyanophenyl)-4-hydroxypyrimidin-2-yl]amino}ethyl)carboxamide,4.82 g (13.5 mmol) of N-phenyltrifluorosulfonimide in 40 mL pyridine wasadded 4.03 g (12.4 mmol) of anhydrous cesium carbonate. The resultingsuspension was stirred at room temperature for 18 h. and thenpartitioned between ethyl acetate and water. The aqueous layer wasextracted thoroughly with ethyl acetate and the combined organics washedwith 5% aqueous hydrochloric acid, water, and brine. Concentration andchromatography (silica gel, ether, Rf=0.40) gavel.50 g of2-({2-[(tert-butoxy)carbonylamino]ethyl}amino)-6-(4-cyanophenyl)pyrimidin-4-yl(trifluoromethyl)sulfonate as a gelatinous solid.2-({2-[(tert-butoxy)carbonylamino]ethyl}amino)-6-(4-cyanophenyl)pyrimidin-4-yl(trifluoromethyl)sulfonate (134 mg, 0.275 mmol) was dissolved in a 2 Msolution of imidazole in N-methylpyrrolidinone and the resultingsolution heated to 90° C. for 18 h. The solution was cooled, water addedand the suspension extracted thoroughly with ethyl acetate.Concentration of the combined organic layers gave a crude solid whichwas chromatographed (silica gel, 0.5% ammonium hydroxide/5%methanol/methylene chloride) to afford 66 mg of(tert-butoxy)-N-(2-{[4-(4-cyanophenyl)-6-imidazolyl-4-ylpyrimidin-2-yl]amino]ethyl)carboxamideas a off white solid.

Removal of the tert-butoxy protecting group was accomplished in thefollowing manner: the(tert-butoxy)-N-(2-{[4-(4-cyanophenyl)-6-imidazolyl-4-ylpyrimidin-2-yl]amino}ethyl)carboxamideproduced as described above was dissolved in anhydrous trifluoroaceticacid (2 mL) and stirred at room temperature for 2 h. Concentration gave47.8 mg of4-{2-[(2-aminoethyl)amino]-6-imidazolyl-4-ylpyrimidin-4-yl}benzene-carbonitrileas its (2×) trifluoroacetate salt.

The4-{2-[(2-aminoethyl)amino]-6-imidazolyl-4-ylpyrimidin-4-yl}benzene-carbonitriletrifluoroacetate produced as described above (48 mg, 0.074 mmol) wasdissolved in 0.5 mL acetonitrile and 2-chloro-5-nitropyridine (12 mg,0.074 m-mol) added. The mixture was heated to 80° C. for 18 h and thenconcentrated. Chromatography of the residue (silica gel, 5%methanol/methylene chloride) afforded 5.1 mg (15%) of4-[6-imidazolyl-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]benzenecarbonitrileas a light yellow solid.

HPLC [Method AZ-S]7.25 min (100%), 1HNMR;

MS: (m+H)/z, 428.

Example 91 Preparation of4-[6-morpholin-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]benzenecarbonitrile

2-({2-[(tert-butoxy)carbonylamino]ethyl}amino)-6-(4-cyanophenyl)pyrimidin-4-yl(trifluoromethyl)sulfonate (134 mg, 0.275 mmol) was dissolved in a 2 Msolution of morpholine in acetonitrile and the resulting solution heatedto 90° C. for 18h. The solution was cooled, water added and thesuspension extracted thoroughly with ethyl acetate. Concentration of thecombined organic layers gave a crude solid which was chromatographed(silica gel, 2% methanol/methylene chloride) to afford 234 mg of(tert-butoxy)-N-(2-{[4-(4-cyanophenyl)-6-morpholin-4-ylpyrimidin-2-yl]amino}ethyl)carboxamideas a off white solid. Removal of the tert-butoxy protecting group wasaccomplished in the following manner: the(tert-butoxy)-N-(2-{[4-(4-cyanophenyl)-6-morpholin-4-ylpyrimidin-2-yl]amino}ethyl)carboxamideproduced as described above was dissolved in anhydrous hydrogen chloridein dioxane (2 mL of a 2M solution) and stirred at room temperature for 2h. Concentration gave 134 mg of4-{2-[(2-aminoethyl)amino]-6-imidazolyl-4-ylpyrimidin-4-yl}benzenecarbonitrileas its (2x) hydrochloride salt.

The4-{2-[(2-aminoethyl)amino]-6-morpholin-4-ylpyrimidin-4-yl}benzene-carbonitriletrifluoroacetate produced above (50 mg, 0.14 mmol) was dissolved in 0.25mL acetonitrile and 2-chloro-5-nitropyridine (22 mg, 0.074 mmol) added.The mixture was heated to 80° C. for 18 h and then concentrated.Chromatography of the residue (silica gel, 5% methanol/methylenechloride) afforded 5.1 mg (15%) of4-[6-morpholin-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]benzenecarbonitrileas a light yellow solid.

HPLC [Method AZ-S]7.20 min (100%), 1HNMR;

MS: (m+H)/z, 447.

Example 92 Preparation of[5-benzotriazolyl-4-(2,4-dichlorophenyl)pyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino[ethyl}amine

A solution of 1-(2,4-dichlorophenyl)-2-chloroethan-1-one (10.0 g, 44.63mmol), benzotriazole (10.6 g, 89.3 mmol) and triethylamine (9.0 g, 89.3mmol) in 140 mL acetonitrile was refluxed for 18h. The acetonitrile wasremoved in vacuo, and the residue dissolved in ethyl acetate. Water wasadded and the water layer extracted twice with ethyl acetate. Thecombined organics were washed with brine, dried and concentrated to give13.2 g of a light brown solid. Recrystallization from ethyl acetate gave4.8 g (35.%) of a colorless solid identified as2-benzotriazolyl-1-(2,4-dichlorophenyl)ethan-1-one.

2-benzotriazolyl-1-(2,4-dichlorophenyl)ethan-1-one was dissolved indimethyl-formamide dimethyl acetal (5 mL) and the solution refluxed for8 h. Evaporation of the solvent gave2-benzotriazolyl-1-(2,4-dichlorophenyl)-3-(dimethylamino)prop-2-en-1-oneas an air sensitive red solid which was used in the next step withoutfurther purification.

2-benzotriazolyl-1-(2,4-dichlorophenyl)-3-(dimethylamino)prop-2-en-1-onewas dissolved in 64 mL N-methylpyrrolidinone and 6.82 g (20.9 mmol) ofcesium carbonate and 4.19 g (16.12 mmol) ofamino{2-[(5-nitro(2-pyridyl))amino]ethyl}carboxamidine hydrochlorideadded. The resulting suspension was heated at 100° C. for 1I8h. Thereaction was then cooled and partitioned between ethyl acetate andwater. The aqueous layer was extracted with ethyl acetate, and thecombined organic layers washed twice with water and once with brine.Concentration of the combined organic layers gave a yellow solid whichwas recrystallized from 5% ethyl acetate/methanol to afford 4.25 g (65%)of[5-benzotriazolyl-4-(2,4-dichlorophenyl)pyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amineas a light yellow solid.

HPLC [Method AZ-S]11.56 min (100%), 1HNMR;

MS: (m+H)/z, 522.

Example 93 Preparation of[4-(2,4-dichlorophenyl)-5-piperazinylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine

1. Preparation of tert-butyl4-[2-(2,4-dichlorophenyl)-2-oxoethyl]piperazinecarboxylate

1 mmol of 2,4-dichlorophenacyl chloride in DMF was added drop wise to 1.mmol of tert-butyl piperazinecarboxylateand 1.2 mmol Cs₂CO₃ in DMF atroom temperature for fourteen hours. The reaction mixture wasconcentrated in vacuo and diluted with water and ethyl acetate. Thesolution was extracted three times with ethyl acetate, dried over sodiumsulfate, and purified by column chromatography eluting with 50%ethylacetate and 50% hexane to obtain tert-butyl4-[2-(2,4-dichlorophenyl)-2-oxoethyl]piperazinecarboxylate.

2. Preparation of tert-butyl4-{2-(2,4-dichlorophenyl)-1-[(dimethylamino)methylene]-2-oxoethyl}piperazinecarboxylate

1 mmol of tert-butyl4-[2-(2,4-dichlorophenyl)-2-oxoethyl]piperazinecarboxylate was heated to80° C. in neat N,N-dimethylformamidedimethyl acetal for six hours. Thereaction mixture was concentrated in vacuo and purified by triturationwith diethyl ether to obtain tert-butyl4-{2-(2,4-dichlorophenyl)-1-[(dimethylamino)methylene]-2-oxo-ethyl}piperazinecarboxylate.

3. Preparation of tert-butyl4-[4-(2,4-dichlorophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-5-yl]piperazinecarboxylate

1 mmol of tert-butyl4-{2-(2,4-dichlorophenyl)-1-[(dimethylamino)methylene]-2-oxoethyl}piperazinecarboxylate,1 mmol of amino{2-[(5-nitro(2-pyridyl))amino]ethyl}carboxamidine, and 3MMol of Cs₂CO₃ was dissolved in DMF and heated to 90° C. for fourteenhours. The reaction mixture was concentrated in vacuo and diluted withwater and ethyl acetate. The solution was extracted three times withethyl acetate, dried over sodium sulfate, and purified by columnchromatography eluting with 5-10% methanol/methylene/chloride to obtaintert-butyl4-[4-(2,4-dichlorophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-5-yl]piperazinecarboxylate.

4. Preparation of[4-(2,4-dichlorophenyl)-5-piperazinylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine

1 mmol of tert-butyl4-[4-(2,4-dichlorophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-5-yl]piperazinecarboxylateis heated to 60° C. 3M HCl in MeOH for one hour and concentrated invacuo to obtain[4-(2,4-dichlorophenyl)-5-piperazinylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC: 7.683 min. (100% purity)

MS: MH⁺=489.1

Example 94 Preparation of1-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]-4-methylpiperazine-2,6-dione

1. Preparation of2-[2-(2,4-dichlorophenyl)-2-oxoethyl]isoindoline-1,3-dione

1 mmol of 2,4-dichlorophenacyl chloride in DMF was added drop wise to 2mmol of phthalimide and 2 mmol of Cs₂CO₃ in DMF at room temperature forfourteen hours and then purified by trituration with diethyl ether toobtain 2-[2-(2,4-dichlorophenyl)-2-oxoethyl]isoindoline-1,3-dione.

2. Preparation of2-{2-(2,4-dichlorophenyl)-1-[(dimethylamino)methylene]-2-oxoethyl}isoindoline-1,3-dione

1 mmol of 2-[2-(2,4-dichlorophenyl)-2-oxoethyl]isoindoline-1,3-dione washeated to 80° C. in N,N-dimethylformamidedimethyl acetal for six hours.The reaction mixture was concentrated in vacuo and purified bytrituration with diethyl ether to obtain2-{2-(2,4-dichlorophenyl)-1-[(dimethylamino)methylene]-2-oxoethyl}isoindoline-1,3-dione.

3. Preparation of2-{N-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]carbamoyl}benzoicacid

1 mmol of2-{2-(2,4-dichlorophenyl)-1-[(dimethylamino)methylene]-2-oxoethyl}isoindoline-1,3-dione,1 mmol ofamino{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}carboxamidine, and 3mmol of Cs₂CO₃ were dissolved in DMF and heated to 90° C. for fourteenhours. The reaction mixture was concentrated in vacuo and diluted withwater and ethyl acetate. The solution was extracted three times withethyl acetate and dried over sodium sulfate to obtain2-{N-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]carbamoyl}benzoicacid.

4. Preparation of2-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]isoindoline-1,3-dione

1 mmol of2-{N-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]carbamoyl}benzoicacid was heated to 120° C. in acetic acid for four hours and thenconcentrated in vacuo to obtain2-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]isoindoline-1,3-dione.

5. Preparation of[5-amino-4-(2,4-dichlorophenyl)pyrimidin-2-yl]{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amine

1 mmol of2-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]isoindoline-1,3-dioneand 20 mmol of hydrazine were stirred in ethanol at 75° C. for two hoursand then purified by column chromatography eluting with 5-10%methanol/methylene chloride to obtain[5-amino-4-(2,4-dichlorophenyl)pyrimidin-2-yl]{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amine.

6. Preparation of1-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]-4-methylpiperazine-2,6-dione

1 mmol of[5-amino-4-(2,4-dichlorophenyl)pyrimidin-2-yl]{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amine,and 2 mmol of methyliminodiacetic acid, 4 mmol of HBTU, and 5 mmol ofN,N-diisopropylethylamine were added to solution and stirred for sixhours at room temperature. The reaction mixture was concentrated invacuo and diluted with water and ethyl acetate. The solution wasextracted three times with ethyl acetate, dried over sodium sulfate, andpurified by column chromatography eluting with 5-10% methanol/methylenechloride to obtain1-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]-4-methylpiperazine-2,6-dione.

HPLC: 7.560min. (99% purity)

MS: MH⁺=546.1

Example 95 Preparation of1-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]-3-morpholin-4-ylpyrrolidine-2,5-dione

1. Preparation of2-[2-(2,4-dichlorophenyl)-2-oxoethyl]isoindoline-1,3-dione

1 mmol of 2,4-dichlorophenacyl chloride in DMF was added drop wise to 2mmol of phthalimide and 2 mmol of Cs₂CO₃ in DMF at room temperature forfourteen hours and then purified by trituration with diethyl ether toobtain 2-[2-(2,4-dichlorophenyl)-2-oxoethyl]isoindoline-1,3-dione.

2. Preparation of2-{2-(2,4-dichlorophenyl)-1-[(dimethylamino)methylene]-2-oxoethyl}isoindoline-1,3-dione

1 mmol of 2-[2-(2,4-dichlorophenyl)-2-oxoethyl]isoindoline-1,3-dione washeated to 80° C. in N,N-dimethylformamidedimethyl acetal for six hours.The reaction mixture was concentrated in vacuo and purified bytrituration with diethyl ether to obtain2-{2-(2,4-dichlorophenyl)-1-[(dimethylamino)methylene]-2-oxoethyl}isoindoline-1,3-dione.

3. Preparation of2-{N-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]carbamoyl}benzoicacid

1 mmol of2-{2-(2,4-dichlorophenyl)-1-[(dimethylamino)methylene]-2-oxoethyl}isoindoline-1,3-dione,1 mmol ofamino{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}carboxamidine, and 3mmol of Cs₂CO₃ were dissolved in DMF and heated to 90° C. for fourteenhours. The reaction mixture was concentrated in vacuo and diluted withwater and ethyl acetate. The solution was extracted three times withethyl acetate and dried over sodium sulfate to obtain2-{N-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]carbamoyl}benzoicacid.

4. Preparation of2-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]isoindoline-1,3-dione

1 mmol of2-{N-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]carbamoyl}benzoicacid is heated to 120° C. in acetic acid for four hours and thenconcentrated in vacuo to obtain2-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]isoindoline-1,3-dione.

5. Preparation of[5-amino-4-(2,4-dichlorophenyl)pyrimidin-2-yl]{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amine

1 mmol of2-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]isoindoline-1,3-dioneand 20 mmol of hydrazine were stirred in ethanol at 75° C. for twohours. The reaction was concentrated and purified by columnchromatography 5-10% methanol/methylene chloride to obtain[5-amino-4-(2,4-dichlorophenyl)pyrimidin-2-yl]{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amine.

5. Preparation of1-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]-3-pyrroline-2,5-dione

1 mmol of[5-amino-4-(2,4-dichlorophenyl)pyrimidin-2-yl]{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amine,and 2 mmol of malaic anhydride are stirred at room temperature for fourhours. 2 mmol of HBTU, and 3 mmol of N,N-diisopropylethylamine wereadded to solution and left for six hours at room temperature. Thereaction mixture was concentrated in vacuo and diluted with water andethyl acetate. The solution was extracted three times with ethylacetate, dried over sodium sulfate, and purified by columnchromatography eluting with 5-10% methanol/methylene chloride to obtain1-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]-3-pyrroline-2,5-dione.

6. Preparation of1-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]-3-morpholin-4-ylpyrrolidine-2,5-dione

Large excess of morpholine was added to clean fractions of1-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]-3-pyrroline-2,5-dioneconcentrated in vacuo, and purified by column chromatography elutingwith 5-10% methanol/methylene chloride to obtain1-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]-3-morpholin-4-ylpyrrolidine-2,5-dione.

HPLC: 8.133 min. (100% purity)

MS: MH⁺=602.2

Example 96 Preparation of1-[4-(2,4-dichlorophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-5-yl]-4-methylpiperazine-2,6-dione

1. Preparation of2-[2-(2,4-dichlorophenyl)-2-oxoethyl]isoindoline-1,3-dione

1 mmol of 2,4-dichlorophenacyl chloride in DMF was added drop wise to 2mmol of phthalimide and 2 mmol of Cs₂CO₃ in DMF at room temperature forfourteen hours and then purified by trituration with diethyl ether toobtain 2-[2-(2,4-dichlorophenyl)-2-oxoethyl]isoindoline-1,3-dione.

2. Preparation of2-{2-(2,4-dichlorophenyl)-1-[(dimethylamino)methylene]-2-oxoethyl}isoindoline-1,3-dione

1 mmol of 2-[2-(2,4-dichlorophenyl)-2-oxoethyl]isoindoline-1,3-dione isheated to 80° C. in neat N,N-dimethylformamidedimethyl acetal for sixhours. The reaction mixture was concentrated in vacuo and purified bytrituration with diethyl ether to obtain2-{2-(2,4-dichlorophenyl)-1-[(dimethylamino)methylene]-2-oxoethyl}isoindoline-1,3-dione.

3. Preparation of2-{N-[4-(2,4-dichlorophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-5-yl]carbamoyl}benzoicacid

1 mmol of2-{2-(2,4-dichlorophenyl)-1-[(dimethylamino)methylene]-2-oxoethyl}isoindoline-1,3-dione,1 mmol of amino {2-[(5-nitro(2-pyridyl))amino]ethyl}carboxamidine, and 3mmol of Cs₂CO₃ were dissolved in DMF and heated to 90° C. for fourteenhours. The reaction mixture was concentrated in vacuo and diluted withwater and ethyl acetate. The solution was extracted three times withethyl acetate and dried over sodium sulfate to obtain2-{N-[4-(2,4-dichlorophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-5-yl]carbamoyl}benzoicacid.

4. Preparation of 2-[4-(2,4-dichlorophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-5-yl]isoindoline-1,3-dione

1 mmol of2-{N-[4-(2,4-dichlorophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-5-yl]carbamoyl}benzoicacid was heated to 120° C. in acetic acid for four hours and thenconcentrated in vacuo to obtain2-[4-(2,4-dichlorophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-5-yl]isoindoline-1,3-dione.

5. Preparation of[5-amino-4-(2,4-dichlorophenyl)pyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine

1 mmol of2-[4-(2,4-dichlorophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-5-yl]isoindoline-1,3-dioneand 20 mmol of hydrazine were stirred in ethanol at 75° C. for two hoursand then purified by column chromatography eluting with 5-10%methanol/methylene chloride to obtain[5-amino-4-(2,4-dichlorophenyl)pyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine.

6. Preparation of 1-[4-(2,4-dichlorophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl{amino)pyrimidin-5-yl]-4-methylpiperazine-2,6-dione

1 mmol of[5-amino-4-(2,4-dichlorophenyl)pyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine,and 2 mmol of methyliminodiacetic acid, 4 mmol of HBTU, and 5 mmol ofN,N-diisopropylethylamine were added to solution and stirred for sixhours at room temperature. The reaction mixture was concentrated invacuo and diluted with water and ethyl acetate. The solution wasextracted three times with ethyl acetate,dried over sodium sulfate,purified by column chromatography eluting with 5-10% methanol/methylenechloride to obtain1-[4-(2,4-dichlorophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-5-yl]-4-methylpiperazine-2,6-dione.

HPLC: 12.850 min. (100% purity)

MS: MH⁺=531.2

Example 97 Preparation of1-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]-3-(dimethylamino)pyrrolidine-2,5-dione

1. Preparation of2-[2-(2,4-dichlorophenyl)-2-oxoethyl]isoindoline-1,3-dione

1 mmol of 2,4-dichlorophenacyl chloride in DMF was added drop wise to 2mmol of phthalimide and 2 mmol of Cs₂CO₃ in DMF at room temperature forfourteen hours and the purified by trituration with diethyl ether toobtain 2-[2-(2,4-dichlorophenyl)-2-oxoethyl]isoindoline-1,3-dione.

2. Preparation of2-{2-(2,4-dichlorophenyl)-1-(dimethylamino)methylene]-2-oxoethyl}isoindoline-1,3-dione

1 mmol of 2-[2-(2,4-dichlorophenyl)-2-oxoethyl]isoindoline-1,3-dione washeated to 80° C. in neat N,N-dimethylformatidedimethyl acetal for sixhours. The reaction mixture was concentrated in vano and purified bytrituration with diethyl ether to obtain2-{2-(2,4-dichlorophenyl)-1-[(dimethylamino)methylene]-2-oxoethyl}isoindoline-1,3-dione.

3. Preparation of2-{N-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]carbamoyl}benzoicacid

1 mmol of2-{2-(2,4-dichlorophenyl)-1-[(dimethylamino)methylene]-2-oxoethyl}isoindoline-1,3-dione,1 mm of amino{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}carboxamnidine,and 3 mmol of Cs₂CO₃ were dissolved in DMF and heated to 90° C. forfourteen hours. The reaction mixture was concentrated in vacuo anddiluted with water and ethyl acetate. The solution was extracted threetimes with ethyl acetate and dried over sodium sufate to obtain2-{N-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]carbamoyl}benzoicacid.

4. Preparation of2-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]isoindoline-1,3-dione

1 mmol of2-{N-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl)}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]carbamoyl}benzoicacid was heated to 120° C. in acetic acid for four hours and thenconcentrated in vacuo to obtain2-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]isoindoline-1,3-dione.

5. Preparation of[5-amino-4-(2,4-dichlorophenyl)pyrimidin-2-yl]{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amine

1 mmol of2-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]isoindoline-1,3-dioneand 20 mmol of hydrazine were stirred in ethanol at 75° C. for two hoursand then purified by column chromatography eluting with 5-10%methanol/methylene chloride to obtain[5-amino-4-(2,4-dichlorophenyl)pyrimidin-2-yl]{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amine.

6. Preparation of1-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]-3-pyrroline-2,5-dione

1 mmol of[5-amino-4-(2,4-dichlorophenyl)pyrimidin-2-yl]{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amine,and 2 mmol of malaic anhydride were stirred at room temperature for fourhours. 2 mmol of HBTU, and 3mmol of N,N-diisopropylethylamine were addedto the solution and left for six hours at room temperature. The reactionmixture was concentrated in vacuo and diluted with water and ethylacetate. The solution was extracted three times with ethyl acetate,dried over sodium sulfate, and purified by column chromatography elutingwith 5-10% methanol/methylene chloride to obtain1-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]-3-pyrroline-2,5-dione.

7. Preparation of1-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]-3-(dimethylamino)pyrrolidine-2,5-dione

A large excess of dimethylamine was added to clean fractions of1-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]-3-pyrroline-2,5-dioneconcentrated in vacuo.

HPLC: 5.2l5 min. (95% purity)

MS: MH⁺=560.2

Example 98 Preparation of{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[6-(2,4-dichlorophenyl)-5-imidazolyl(2-pyridyl)]amine

1. Preparation of 1-(2,4-dichlorophenyl)-2-imidazolylethan-1-one.

A stirred solution of 1-(2,4-dichlorophenyl)-2-chloroethan-1-one (0.95M) and imidazole (2.68 M) in CH₃CN (500 ml) were heated for 14-16 hoursat 75° C. The product was stripped of solvent under reduced pressure.The residue was diluted with dichloromethane (1 L) and water (400 ml),and the mixture was stirred for 30 min. After filtering off a solidimpurity, the aqueous layer was removed and discarded. The organic layerwas washed sequentially with water (300 ml), sat. aq. NaHCO₃ (300 ml),water (300 ml), brine (200 ml), dried with Na₂SO₄, filtered, andconcentrated under reduced pressure. The reddish black oil was driedovernight in vacuo giving 1-(2,4-dichlorophenyl)-2-imidazolylethan-1-onein 90% yield.

2. Preparation of ethyl5-(2,4-dichlorophenyl)-4-imidazolyl-5-oxopentanoate.

1-(2,4-dichlorophenyl)-2-imidazolylethan-1-one (0.80 M) was dissolvedwith heating as necessary in a stirred mixture of THF (250 ml) and abs.ethanol (250 ml). After cooling to room temperature, potassium hydroxide(0.20 M) was added followed by a dropwise addition of ethyl acrylate(45.8 ml) via addition funnel over 10 min. A room temperature water bathwas used to cool the exothermic reaction for the first 30 min. Afterstirring for 14-16 hours, the reaction was neutralized with gla. aceticacid (10 ml) and concentrated under reduced pressure. The residual darkreddish black slurry was dissolved in abs. ethanol (100 ml) andconcentrated again under reduced pressure giving ethyl5-(2,4-dichlorophenyl)-4-imidazolyl-5-oxopentanoate (2) in 108% yield.The crude material is contaminated with potassium salts and is usedwithout further purification.

3. Preparation of6-(2,4-dichlorophenyl)-5-imidazolyl-1,3,4-trihydropyridin-2-one.

A mixture of ethyl 5-(2,4-dichlorophenyl)-4-imidazolyl-5-oxopentanoate(0.51 M) was dissolved in gla. acetic acid (245 ml), toluene (135 ml),and abs. ethanol (405 ml). Ammonium acetate (3.07 M) and flame dried 4Apowder molecular sieves (145 g) were added to the stirred solution. Theresulting mixture was stirred for 44-46 hours at 90-95° C. under argon.After 24 hours of heating, additional reagents were added includingammonium acetate (0.51 M), acetic acid (41 ml), and flame dried 4Apowder molecular sieves (24 g). On cooling methanol (200 ml) was addedwith stirring for 15 minutes. The sieves were filtered and washed withmethanol (2×150 ml). The filtrate was concentrated under reducepressure. To the crude material was added dichloromethane (1.5 L). Theorganic layer was then washed until basic, with 5-10% sodium hydroxidesolution (3×500 ml). The organic layer was then washed with distilledwater (3×400 ml), saturated sodium chloride solution (300 ml), driedwith sodium sulfate, filtered, and concentrated under reduce pressure.The reddish orange solid was dried in vacuo giving 102 g of crudeproduct. A solution of 2% methanol in ethyl acetate (90-110 ml) wasadded to the crude product. The resulting solid was collected byfiltration and washed with ethyl acetate (2×100 ml). The off white solidwas dried in vacuo to give6-(2,4-dichlorophenyl)-5-imidazolyl-1,3,4-trihydropyridin-2-one in 60%yield.

4. Preparation of 6-(2,4-dichlorophenyl)-5-imidazolylhydropyridin-2-one.

A mixture of6-(2,4-dichlorophenyl)-5-imidazolyl-1,3,4-trihydropyridin-2-one (0.21M), selenium(IV) oxide (0.63), and gla. acetic acid (400 ml) was stirredfor 10 hours at 105-110° C. under argon. The acetic acid was removedunder reduced pressure. Methanol (500 ml) was added. After mixing, thesolution was filtered to remove selenium residue. To the methanolicsolution, lead(II) acetate trihydrate (99 g) and distilled water (50 ml)were added and stirred for 1 hour. The mixture was filtered through acelite plug (0.25-0.5 inches). The solution was concentrated underreduced pressure. The crude material was purified by a silica gelcolumn. The product was eluted with ethyl acetate and a slowlyincreasing gradient of methanol reaching a final concentration of 8%.The proper fractions were concentrated under reduced pressure and driedin vacuo. The solid was further purified by trituration with a smallvolume of 1:1 methanol and ethyl acetate. The off-white solid was driedin vacuo to give 6-(2,4-dichlorophenyl)-5-imidazolylhydropyridin-2-onein 68% yield.

5. Preparation of [2-(2,4-dichlorophenyl)-6-chloro-3-pyridyl]imidazole.

To the dry solid 6-(2,4-dichlorophenyl)-5-imidazolylhydropyridin-2-one(2.40 M) was added N,N-dimethylacetamide (6 drops) followed byphosphorous oxychloride (20 ml). The reaction mixture was stirred for18-20 hours at 105-110° C. under argon. The phosphorous oxychloride wasremoved under reduced pressure. The crude product was taken up indichloromethane (75 ml), and the solvent was removed under vacuum. Thesticky solid was dried to a free flowing solid in vacuo 3-4 hour giving[2-(2,4-dichlorophenyl)-6-chloro-3-pyridyl]imidazole in 167% yield. Thecrude material is contaminated with phosphorous residue and is used without further purification.

6. Preparation of(2-aminoethyl)[6-(2,4-dichlorophenyl)-5-imidazolyl(2-pyridyl)]amine.

The crude material above,[2-(2,4-dichlorophenyl)-6-chloro-3-pyridyl]imidazole was cooled underargon to −78° C. with dry ice acetone. With argon continually flushingthrough the system, ethylene diamine (200 ml) was added very carefullymonitoring the release of gas and heat. After completing the addition,the reaction mixture was stirred for 5-6 hours at 105-110° C. underargon. Upon cooling, the ethylene diamine was removed under reducedpressure and dried in vacuo for 2-3 hours. To the residual material wasadded acetonitrile (100 ml) and saturated sodium bicarbonate solution(250-300 ml). To the stirred mixture above, sodium bicarbonate solid wasadded until the mixture was fully saturated. After 30 min., acetonitrile(300-350 ml) was added to the saturated aqueous solution. The mixturewas stirred and the layers were separated saving both aqueous andorganic portions. The aqueous layer was extracted with acetonitrile(4×250 ml). The organic layers were combined and washed with saturatedsodium chloride solution (100 ml), dried with sodium sulfate, filtered,concentrated, and dried in vacuo. The residue was dissolved in methanol(70-90 ml). The mixture was filtered to remove residual salt, andconcentrated under reduced pressure. The off-white solid was dried invacuo giving(2-aminoethyl)[6-(2,4-dichlorophenyl)-5-imidazolyl(2-pyridyl)]amine in97% yield based on the initial amount of6-(2,4-dichlorophenyl)-5-imidazolylhydropyridin-2-one used. The materialcontains minimal amounts of salt and was used without furtherpurification.

7. Preparation of{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[6-(2,4-dichlorophenyl)-5-imidazolyl(2-pyridyl)]amine.

A mixture of(2-aminoethyl)[6-(2,4-dichlorophenyl)-5-imidazolyl(2-pyridyl)]amine(0.14 M), 6-chloro-3-nitro-2-pyridylamine (0.14 M),N-N-dimethylacetamide (40 ml), and Hünig's base (2.5 ml) was stirred for12 hours at 85-90° C. under argon. Ethylene diamine (3.8 ml) was addedto remove the unreacted chloropyridine, and the mixture was stirred foran additional 0.75-1 hour at 85-90° C. under argon. After cooling, thereaction was diluted with ethyl acetate (500-600 ml), extracted withsaturated sodium bicarbonate (4×200 ml), distilled water (3×150 ml),saturated sodium chloride (150 ml). The organic layer was dried withsodium sulfate briefly (2-3 minutes) so as not to initiate precipitationof the product. The organic layer was filtered, concentrated, and driedin vacuo. To cause precipitation of the product, a minimal amount ofmethanol (3-5 ml) was added followed with the addition of an equalvolume of ethyl acetate. The mixture was allowed to age for 2-3 hours.The solid was collected by filtration, washed with minimal 1:1methanol/ethyl acetate (5 ml), and a finally 100% ethyl acetate (2×10ml). The yellow solid was dried in vacuo to give{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[6-(2,4-dichlorophenyl)-5-imidazolyl(2-pyridyl)]aminein61% yield.

HPLC: 19.8 min (>95% purity)

MS: M+H=485 (C₂₁H₁₈C₁₂N₈O₂+H=485)

Example 99 Preparation of6-[(2-{[4-(2,4-dichlorophenyl)-5-(4-methylimidazol-2-yl)pyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carbonitrile

6-[(2-{[4-(2,4-dichlorophenyl)-5-(4-methylimidazol-2-yl)pyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carbonitrilewas prepared using the general method for[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}aminewith the exceptions noted below.

1. Preparation of1-(2,4-dichlorophenyl)-2-(4-methylimidazol-2-yl)ethan-1-one.

A solution of 2,4-dichlorobenzoyl chloride (7.24 M) in dichloromethane(25 ml) was added dropwise over 20 minutes to a stirred solution of2,4-dimethylimidazole (0.80 M) in dichloromethane (75 ml) andN,N-diisopropylethylamine (Hünig's base) (34 ml). The reaction mixturewas cooled during the addition using a water bath. The reaction mixturewas then heated to reflux for 5 hours. The reaction can turn a darkercolor. The product was stripped of solvent under reduced pressure, andthe resulting solid was dried in vacuo for one hour.

To the dry solid (described above) was added a solution (2:1 v/v, 120ml) of gla. acetic acid and aq. con. HCl. The mixture was then stirredat reflux for ca. 90 min. The acetic acid was removed via rotaryevaporator. Upon cooling, distilled water (200 ml) and toluene (100 ml)were added to the solid residue, which was vigorously stirred for 30min. The solids were filtered, rinsed with 50 ml distilled water, anddiscarded. The filtrate was transferred to a separatory funnel. Afterthe organic layer was discarded, the aqueous layer was washed withtoluene (2×100 ml). The aqueous layer was transferred to a large beaker(2 L) and diluted with isopropyl ether (50 ml). The stirred mixture wasbasified (pH 7-8) by careful addition of sodium bicarbonate which leadsto the formation of a sticky white solid. Dichloromethane (200 ml) wasadded and stirring continued for 10 min. The organic layer was separatedand the aqueous layer was again extracted with dichloromethane (100 ml).The organic layers were combined and washed with sat. aq. NaHCO₃ (100ml), distilled water (100 ml), brine (100 ml), dried with Na₂SO₄,filtered, concentrated, and dried in vacuo giving1-(2,4-dichlorophenyl)-2-(4-methylimidazol-2-yl)ethan-1-one in 46%yield.

2. Preparation of(2Z)-1-(2,4-dichlorophenyl)-3-(dimethylamino)-2-(4-methylimidazol-2-yl)prop-2-en-1-one.

A mixture of 1-(2,4-dichlorophenyl)-2-(4-methylimidazol-2-yl)ethan-1-one(0.33 M) and N,N-dimethylformamide-dimethyl acetal (DMFDMA) (25 ml) wasstirred for 2.5 h at 70-75° C. The DMFDMA was then removed under reducedpressure and dried under high vacuum for several hours giving a lightorange solid in quantitative yield. The enaminone product(2Z)-1-(2,4-dichlorophenyl)-3-(dimethylamino)-2-(4-methylimidazol-2-yl)prop-2-en-1-onewas typically used without further purification.

3. Preparation of 6-[(2-aminoethyl)amino]pyridine-3-carbonitrile.

A mixture of 2-chloro-5-cyanopyridine (0.60 M) in acetonitrile (120 ml)and ethylene diamine (85 ml) were stirred overnight (ca. 16 h) at 75-80°C. under argon. The ethylene diamine was removed under reduced pressureand then dried in vacuo for 2-3 h. The residual solution was basifiedwith 1M sodium hydroxide solution (˜100 ml). The aqueous solution wassaturated with sodium chloride and extracted with a solution of 95%ethyl acetate and 5% methanol (3×150 ml) and with a solution of 95%acetonitrile and 5% methanol (3×150 ml). The organic extracts werecombined and extracted with a saturated sodium chloride solution (2×70ml). The organic layer was dried with sodium sulfate, filtered, andconcentrated under reduced pressure. The crude white to tan solid wastriturated with ether (2×50 ml) and dried overnight in vacuo resultingin 78% yield of 6-[(2-aminoethyl)amino]pyridine-3-carbonitrile.

4. Preparation of amino{2-[(5-cyano(2-pyridyl))amino]ethyl}carboxamidine, hydrochloride.

A mixture of 6-[(2-aminoethyl)amino]pyridine-3-carbonitrile (0.47 M),1H-pyrazole-1-carboxamidine hydrochloride (0.47 M), and acetonitrile(120 ml) were stirred ca. 24 h at 75-80° C. Upon cooling, a precipitatewas collected by filtration. The white solid was washed thoroughly withacetonitrile (2×100 ml), ethyl ether (3×100 ml), and dried overnight invacuo giving amino{2-[(5-cyano(2-pyridyl))amino]ethyl}carboxamidine asthe HCl salt in 82% yield.

5. Preparation of6-[(2-{[4-(2,4-dichlorophenyl)-5-(4-methylimidazol-2-yl)pyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carbonitrile.

A solution of sodium ethoxide (0.58 M) dissolved in abs. ethanol (15 ml)was added to a stirred mixture of(2Z)-1-(2,4-dichlorophenyl)-3-(dimethylamino)-2-(4-methylimidazol-2-yl)prop-2-en-1-one(0.41 M), amino{2-[(5-cyano(2-pyridyl)) amino]ethyl}carboxamidine,hydrochloride (0.43 M), and abs. ethanol (20 ml). The reaction was thenheated to 75-80° C. for 2.5 hours. On cooling the reaction was dilutedwith ethyl acetate (400 ml) washed with sat. aq. NaHCO₃ (100 ml),distilled water (2×100 ml), brine (100 ml), dried with Na₂SO₄, filtered,and concentrated. The crude product (˜50% purity) was purified by flashchromatography over silica gel. The column was run starting with 1:1ethyl acetate to hexane, then ethyl acetate which was used until all ofthe fast moving impurities had been removed. The product was eluted with1.5% methanol in ethyl acetate. The column is monitored by TLC using 5%methanol in ethyl acetate as the solvent system. The product has UVactivity in the long wave length region and “glows” blue on theunstained TLC plate. The proper fractions were condensed. The off-whitesolid was dried overnight in vacuo giving6-[(2-{[4-(2,4-dichlorophenyl)-5-(4-methylimidazol-2-yl)pyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carbonitrilein 28% yield.

HPLC: 20.7 min (>99% purity)

MS: M+H=465.3 (C₂₂H₁₈C₁₂N₈+H=465)

Example 100 Preparation of[5-((1E)-1-aza-2-morpholin-4-ylprop-1-enyl)-4-(2,4-dichlorophenyl)pyrimidin-2yl]{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amine

1. Preparation of2-[2-(2,4-dichlorophenyl)-2-oxoethyl]isoindoline-1,3-dione

1 mmol of 2,4-dichlorophenacyl chloride in DMF was added drop wise to 2mmol of phthalimide and 2 mmol of Cs₂CO₃ in DMF at room temperature forfourteen hours and then purified by trituration with diethyl ether toobtain 2-[2-(2,4-dichlorophenyl)-2-oxoethyl]isoindoline-1,3-dione.

2. Preparation of2-{2-(2,4-dichlorophenyl)-1-[(dimethylamino)methylene]-2-oxoethyl}isoindoline-1,3-dione

1 mmol of 2-[2-(2,4-dichlorophenyl)-2-oxoethyl]isoindoline-1,3-dione washeated to 80° C. in N,N-dimethylformamidedimethyl acetal for six hours.The reaction mixture was concentrated in vacuo and purified bytrituration with diethyl ether to obtain2-{2-(2,4-dichlorophenyl)-1-[(dimethylamino)methylene]-2-oxoethyl}isoindoline-1,3-dione.

3. Preparation of2-{N-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]carbamoyl}benzoicacid

1 mmol of2-{2-(2,4-dichlorophenyl)-1-[(dimethylamino)methylene]-2-oxoethyl}isoindoline-1,3-dione,1 mmol ofamino{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}carboxamidine, and 3mmol of Cs₂CO₃ were dissolved in DMF and heated to 90° C. for fourteenhours. The reaction mixture was concentrated in vacuo and diluted withwater and ethyl acetate. The solution was extracted three times withethyl acetate and dried over sodium sulfate to obtain2-{N-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]carbamoyl}benzoicacid.

4. Preparation of2-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]isoindoline-1,3-dione

1 mmol of2-{N-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]carbamoyl}benzoicacid was heated to 120° C. in acetic acid for four hours andconcentrated in vacuo to obtain2-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]isoindoline-1,3-dione.

5. Preparation of[5-amino-4-(2,4-dichlorophenyl)pyrimidin-2-yl]{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amine

1 mmol of2-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]isoindoline-1,3-dioneand 20 mmol of hydrazine were stirred in ethanol at 75° C. for two hoursand purified by column chromatography eluting with 5-10%methanol/methylene chloride to obtain[5-amino-4-(2,4-dichloro-phenyl)pyrimidin-2-yl]{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amine.

6. Preparation ofN-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]acetamide

1 mmol of[5-amino-4-(2,4-dichlorophenyl)pyrimidin-2-yl]{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amineand 1 mmol of acetic anhydride were stirred at room temperature for fourhours in THF. The reaction mixture was concentrated in vacuo and dilutedwith water and ethyl acetate. The solution was extracted three timeswith ethyl acetate, dried over sodium sulfate, and purified by columnchromatography eluting with 5-10% methanol/methylene chloride to obtainN-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]acetamide.

7. Preparation of1-{[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]amino}ethane-1-thione

1 mmol ofN-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]acetamideand 2 mmol of Lawesson's reagent were stirred in 2 ml of DME at 80° C.The reaction mixture was concentrated in vacuo and diluted with waterand ethyl acetate. The solution was extracted three times with ethylacetate, dried over sodium sulfate, and purified by columnchromatography eluting with 5-10% methanol/methylene chloride to obtain1-{[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]amino}ethane-1-thione.

8. Preparation of[5-((1Z)-1-aza-2-morpholin-4-ylprop-1-enyl)-4-(2,4-dichlorophenyl)pyrimidin-2-yl]{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amine

1 mmol of1-{[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]amino}ethane-1-thionewas heated to 90° C. in morpholine and purified by column chromatographyeluting with 5-10% methanol/methylene chloride to obtain [5-((1Z)-1-aza-2-morpholin-4-ylprop-1-enyl)-4-(2,4-dichlorophenyl)pyrimidin-2-yl]{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC: 9.75 min. (100% purity)

MS: MH⁺=546.3

Example 101 Preparation of[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(6-methoxy-5-nitro(2-pyridyl))amino]ethyl}amine

1. Preparation of 6-chloro-2-methoxy-3-nitro-pyridine

To a suspension of sodium hydride (684 mg, 28.49 mmol) in xylene (100ml), methanol (0.98 ml, 25.9 mmol) in xylene (30 ml) was added undernitrogen. The mixture was stirred for 20 min. A solution of2,6-dichloro-3-nitropyridine (5.0g, 25.9mmo) in xylene (100 ml) wasadded to the reaction mixture and stirred at room temperature overnight.Water (50 ml) was added and the organic layer was separated. Theorganics was washed with water (1×50 ml) and brine (2×50 ml), dried, andconcentrated in vacuo. The crude product was purified by flashchromatography (10:1 methylene chloride and acetone) to provide thedesired compound, 6-chloro-2-methoxy-3-nitro-pyridine, as the onlyisomer as light yellow solid (90%).

2. Preparation of[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(6-methoxy-5-nitro(2-pyridyl))amino]ethyl}amine

To a solution of[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-ylethylamine (20 mg,0.04 mmol) in DMF (1 ml), 2-methoxy-3-nitro-6-chloro-pyridine (8.3 mg,0.04 mmol) and diisopropylethyl amine (31 μl, 0.18 mmol) were added. Thereaction mixture was stirred for 12 hours at 80° C. The crude mixturewas concentrated in vacuo and subjected to column chromatography (10%methanol in methylene chloride) to afford[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(6-methoxy-5-nitro(2-pyridyl))amino]ethyl}amineas bright yellow solid.

HPLC: 3.1 min (100% pure)

MS: MH+=501 C21H18C12N8O3=500 g/mol

Example 102 Preparation of[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl]{2-[(6-methoxy-5-nitro(2-pyridyl))amino]ethyl}amine

[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl]{2-[(6-methoxy-5-nitro(2-pyridyl))amino]ethyl}aminewas prepared from[4-(2,4-dichlorophenyl)-5-imidazol-1-ylpyrimidin-2-ylethylamine inaccordance to the procedure described above for the preparation of[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(6-methoxy-5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC: 3.2 min (100% pure)

MS: MH+=501 C21H18C12N8O3=500 g/mol

Example 103 Preparation of6-[(2-{[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amino}ethyl)aminol-3-nitropyridin-2-ol

[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(6-methoxy-5-nitro(2-pyridyl)amino]ethyl}amine(7 mg, 0.01 mmol) was treated with hydrobromic acid (100?1) and aceticacid (100?1) and stirred at 100° C. overnight. The mixture wasconcentrated in vacuo and lyophilized to give6-[(2-{[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amino}ethyl)amino]-3-nitropyridin-2-olas a dark brown solid.

HPLC: 2.7 min (100% pure)

MS: MH+=487 C20H16C12N8O3=486 g/mol

Example 104 Preparation of6-[(2-{[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl]amino}ethyl)amino]-3-nitropyridin-2-ol

6-[(2-{[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl]amino}ethyl)amino]-3-nitropyridin-2-olwas prepared from[4-(2,4-dichlorophenyl)-5-imidazol-1-ylpyrimidin-2-yl]{2-[(6-methoxy-5-nitro(2-pyridyl)amino]ethyl}amine(7mg, 0.01 mmol) by following the same procedure as described for thepreparation of6-[(2-{[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amino}ethyl)amino]-3-nitropyridin-2-ol.

HPLC: 2.46 min (100% pure)

MS: MH+=487 C20H16C12N8O3=486 g/mol

Example 105 Preparation of1-{6-[(2-{[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl]amino}ethyl)amino]-3-pyridyl}ethan-1-one

1-{6-[(2-{[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl]amino}ethyl)amino]-3-pyridyl}ethan-1-onewas prepared from[4-(2,4-dichlorophenyl)-5-imidazol-1-ylpyrimidin-2-yl]ethylamine and1-(6-chloro-3-pyridyl)ethanone (prepared as described in Tetrahedron48:9233 (1992)) in accordance to the procedure described above for thepreparation of[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(6-methoxy-5-nitro(2-pyridyl))amino]ethyl}amine.

Example 106 Preparation of[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl](2-{[5-(iminomethoxymethyl)(2-pyridyl)]amino}ethyl)amineAnd6-[(2-{[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carboxamide

[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-cyano(2-pyridyl)amino]ethyl}amine(20 mg, 0.04 mmol) was treated with saturated solution of methanolicammonia (1 ml) and ammonium chloride (10 mg) and stirred at 60° C.overnight. The crude product was purified by column chromatography (10%methanol in methylene chloride) to provide the title compounds as whitepowder.

HPLC: 2.38 min (100% pure)

MS: MH+=484 C22H20C12N7O=483 g/mol

HPLC: 1.94 min (100% pure)

MS: MH+=469 C22H20C12N7O=468 g/Mol

Example 107 Preparation of{6-[(2-{[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amino}ethyl)amino](3-pyridyl)}iminomethylhydroxylamine

A solution of[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-cyano(2-pyridyl)amino]ethyl}amine(20 mg, 0.04 mmol) in aqueous ethanol(50?l of water in 500 ?l ethanol)was treated with hydoxylamine hydrochloride (3.0 mg, 0.04 mmol) anddiisopropylethyl amine (16 ?l, 0.08 mmol). The reaction mixture wasstirred overnight under reflux, and concentrated in vacuo. The residuewas taken up in ethylacetate and washed with brine and concentrated. Thecrude mixture was subjected to column chromatography (10% methanol inmethylene chloride) togive{6-[(2-{[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amino}ethyl)amino](3-pyridyl)}imino-methylhydroxylamine.

HPLC: 2.16 min (100% pure)

MS: MH+=484 C21H190C12N90O=483 g/mol

Example 108 Preparation of{6-[(2-{[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl]amino}ethyl)amino](3-pyridyl)}iminomethylhydroxylamine

{6-[(2-{[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl]amino}ethyl)amino](3-pyridyl)}iminomethylhydroxylaminewas prepared from[4-(2,4-dichlorophenyl)-5-imidazol-1-ylpyrimidin-2-yl]ethylamine inaccordance to the procedure described for the preparation of{6-[(2-{[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amino}ethyl)amino](3-pyridyl)}iminomethylhydroxylamine.

HPLC: 2.19 min (100% pure)

MS: MH+=484 C21H190C12N9O=483 g/mol

Example 109 Preparation of6-[(2-{[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carboxamidine

A solution of[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-cyano(2-pyridyl)amino]ethyl}amine(20 mg, 0.04 mmol) in saturated solution of methanoic ammonia wasstirred at 60° C. for 48 h. The product was purified from the crudeusing reverse phase column chromatography to give6-[(2-{[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carboxamidine.

HPLC: 2.14 min (100% pure)

MS: MH+=468 C21H19C12N9=467 g/mol

Example 110 Preparation of[4-(2,4-dichlorophenyl)-5-imidazol-1-ylpyrimidin-2-yl]{2-[(4-cyano(2-pyridyl)amino]ethyl}amine

[4-(2,4-dichlorophenyl)-5-imidazol-1-ylpyrimidin-2-yl]{2-[(4-cyano(2-pyridyl)amino]ethyl}aminewas prepared from[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]ethylamine and2-chloro-4-cyanopyridine in accordance to the procedure described abovefor the preparation of[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(6-methoxy-5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC: 2.79 min (100% pure)

MS: MH+=451 C21H16C12N8=450 g/mol

Example 111 Preparation of{6-[(2-{[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amino}ethyl)amino]-3-nitro(2-pyridyl)}dimethylamine

1. Preparation of 6-chloro-2-dimethylamino-3-nitro pyridine

A mixture of 2,6-dichloro-3-nitropyridine (1.9 g, 10 mmole) andpotassium carbonate (1.66 g, 12 mmole) in 30 ml of tetrahydrofuran wasstirred at 0° C. for 5 min. A solution of dimethylamine/tetrahydrofuran(2 M, 6 ml, 12 mmole) was added dropwise to the reaction mixture over 40min. After stirring for 5 min at 0° C., the mixture was warmed to roomtemperature and stirred overnight. The reaction mixture was filtered andthe filtrate was collected and concentrated under reduced pressure. Thecrude product was purified by flash chromatography, eluting with 87%hexane: 13% ethyl acetate to give

6-chloro-2-dimethylamino-3-nitro pyridine (1.05 g).

HPLC: 11.18 min (90% pure)

MS: MH+=202.1 C7H8C1N3O2=201 g/mol

2. Preparation of{6-[(2-{[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amino}ethyl)amino]-3-nitro(2-pyridyl)}dimethylamine

{6-[(2-{[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amino}ethyl)amino]-3-nitro(2-pyridyl)}dimethylaminewas prepared from[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-ylethylamine and6-chloro-2-dimethylamino-3-nitro pyridine in accordance with theprocedure described above for the preparation of[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(6-methoxy-5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC: 11.5 min (85% pure)

MS: MH+=514.2 C22H21C12N9O2=513 g/mol

Example 112 Preparation of{6-[(2-{[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl]amino}ethyl)amino]-3-nitro(2-pyridyl)}dimethylamine

{6-[(2-{[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl]amino}ethyl)amino]-3-nitro(2-pyridyl)}dimethylaminewas prepared from[4-(2,4-dichlorophenyl)-5-imidazol-1-ylpyrimidin-2-ylethylamine and6-chloro-2-dimethylamino-3-nitro pyridine in accordance with theprocedure described above for the preparation of[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(6-methoxy-5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC: 11.6 min (85% pure)

MS: MH+=514.3 C22H21C12N9O2=513 g/mol

Example 113 Preparation of[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl](2-{[6-(methylamino)-5-nitro(2-pyridyl)]amino}ethyl)amine

1. Preparation of 6-chloro-2-methylamino-3-nitro-pyridine

6-chloro-2-methylamino-3-nitro-pyridine was prepared in accordance tothe procedure described above for the preparation of6-chloro-2-dimethylamino-3-nitro pyridine by using solution of methylamine. The crude product was purified by flash chromatography, elutingwith 90% hexane: 10% ethyl acetate to 16 (300 mg).

HPLC: 12.06 min (85% pure)

MS: MH+=188.1 C6H6C1N3O2 187 g/mol

Preparation of[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl](2-{[6-(methylamino)-5-nitro(2-pyridyl)]amino}ethyl)amine

[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl](2-{[6-(methylamino)-5-nitro(2-pyridyl)]amino}ethyl)aminewas prepared from[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-ylethylamine and6-chloro-2-methylamino-3-nitro-pyridine in accordance with the proceduredescribed above for the preparation of[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(6-methoxy-5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC: 11 min (85% pure)

MS: MH+=500.3 C21H19C12N9O2 499 g/mol

Example 114 Preparation of[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl](2-{[6-(methylamino)-5-nitro(2-pyridyl)]amino}ethyl)amine

[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl](2-{[6-(methylamino)-5-nitro(2-pyridyl)]amino}ethyl)aminewas prepared from[4-(2,4-dichlorophenyl)-5-imidazol-1-ylpyrimidin-2-ylethylamine and6-chloro-2-methylamino-3-nitro-pyridine in accordance with the proceduredescribed above for the preparation of[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(6-methoxy-5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC: 11.3 min (85% pure)

MS: MH+=500.3 C21H19C12N9O2 499 g/mol

Example 115 Preparation of 2,6-dichloropyridine-3-carboxamide;2,6-dichloropyridine-3-carbonitrile; and6-chloro-2-methoxypyridine-3-carbonitrile

1. Preparation of 2,6-dichloropyridine-3-carboxamide

A mixture of 2,6-dichloro-3-carboxypyridine (5.73 g, 30 mmole) andN-methylmorpholine (3.6 ml, 33 mmole) in dichloromethane (100 ml) wasstirred in an ice-bath for 5 min. Isopropyl chloroformate (4.28 ml, 33mmole) was added the reaction mixture at 0° C. The reaction mixture wasstirred for 30 min and then bubbled with pure ammonia gas for 2 min. andstirred overnight at room temperature. The crude was concentrated underreduced pressure and sodium bisulfate (0.5 M, 35 ml) was added withstirring. The aqueous solution was extracted with ethyl acetate (3×40ml). The organics was washed with water and brine and dried with sodiumsulfate to give yellow crude product (6.3g, 50% pure).

Preparation of 2,6-dichloropyridine-3-carbonitrile

To a mixture of 2,6-dichloro-3-acetamidopyridine (6.3 g, 55% pure) andpyridine (4.93 ml, 61 mmole) in dichloromethane (100 ml),trifluoroacetic anhydride (4.23 ml, 30 mmole) was added. The reactionmixture was stirred at room temperature overnight and concentrated underreduced pressure. The crude product was purified by flashchromatography, eluting with 85% hexane: 15% ethyl acetate to give apale yellow product (1.77 g, 90%).

HPLC: 10.26 min (90% pure)

MS: MH+=172.9 C6H2C12N2 171.9 g/mol

Preparation of 6-chloro-2-methoxypyridine-3-carbonitrile

6-chloro-2-methoxypyridine-3-carbonitrile was prepared from2,6-dichloro-pyridine-3-carboxamide in accordance with the proceduredescribed above for the preparation of6-chloro-2-methoxy-3-nitro-pyridine.

HPLC: 11.37 min (80% pure)

MS: MH+=169.1 C7H5C1N2O 168 g/mol

Example 116 Preparation of6-[(2-{[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl]amino}ethyl)amino]-2-methoxypyridine-3-carbonitrile

6-[(2-{[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl]amino}ethyl)amino]-2-methoxypyridine-3-carbonitrilewas prepared from[4-(2,4-dichlorophenyl)-5-imidazol-1-ylpyrimidin-2-ylethylamine and6-chloro-2-methoxypyridine-3-carbonitrile in accordance with theprocedure described above for the preparation of[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(6-methoxy-5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC: 11.8 min (85% pure)

MS: MH+=481.2 C22H18C12N8O 480 g/mol

Example 117 Preparation of6-[(2-{[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amino}ethyl)amino]-2-methoxypyridine-3-carbonitrile

6-[(2-{[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amino}ethyl)amino]-2-methoxypyridine-3-carbonitrilewas prepared from[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-ylethylamine and6-chloro-2-methoxypyridine-3-carbonitrile in accordance with theprocedure described above for the preparation of[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(6-methoxy-5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC: 11.37 min (80% pure)

MS: MH+=169.1 C7H5C1N2O 168 g/mol

Example 118 Preparation ofN-{6-[(2-{[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amino}ethyl)amino]-3-nitro-2-pyridyl}acetamide

1. Preparation of N-(6-chloro-3-nitro-2-pyridyl)acetamide

To a suspension of sodium hydride (120 mg, 5 mmole) intetrahydrofuran(10 ml), a solution of 2-amino-3-nitro-6-chloropyridine(870 mg, 5 mmole) in 20 ml of tetrahydrofuran (20 ml) was added. Thereaction mixture was stirred at room temperature for 30 min. A solutionof acetic anhydride (377 μl, 6 mmole) in tetrahydrofuran (10 ml) wasadded and stirred at room temperature overnight. The reaction mixturewas quenched with water and concentrated under reduced pressure. Thecrude product was dissolved in dichloromethane and washed with brine.The organic layer was dried with sodium sulfate and then concentratedunder vacuo. The crude was purified by flash chromatography, eluting 78%hexane: 22% ethyl acetate to give a yellow product (88 mg).

HPLC: 6.36 min and 9.78 min (88% pure)

MS: MH+=215.9 C7H6C1N3O3 215 g/mol

2. Preparation ofN-{6-[(2-{[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amino}ethyl)amino]-3-nitro-2-pyridyl}acetamide

N-{6-[(2-{[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amino}ethyl)amino]-3-nitro-2-pyridyl}acetamidewas prepared from[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-ylethylamine andN-(6-chloro-3-nitro-2-pyridyl)acetamide in accordance with the proceduredescribed above for the preparation of[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(6-methoxy-5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC: 10.6 min (85% pure)

MS: MH+=528.2 C22H19C12N9O3 527 g/mol

Example 119 Preparation ofN-{6-[(2-{[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl]amino}ethyl)amino]-3-nitro-2-pyridyl}acetamide

N-{6-[(2-{[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl]amino}ethyl)amino]-3-nitro-2-pyridyl}acetamidewas prepared from[4-(2,4-dichlorophenyl)-5-imidazol-1-ylpyrimidin-2-ylethylamine andN-(6-chloro-3-nitro-2-pyridyl)acetamide in accordance with the proceduredescribed above for the preparation of[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(6-methoxy-5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC: 10.8 min (85% pure)

MS: MH+=528.2 C22H19C12N9O3 527 g/mol

Example 120 Preparation of{6-[(2-{[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amino}ethyl)amino]-3-nitro(2-pyridyl)}(methylsulfonyl)amine

1. Preparation of (6-chloro-3-nitro(2-pyridyl))(methylsulfonyl)amine

(6-chloro-3-nitro(2-pyridyl))(methylsulfonyl)amine was prepared from2-amino-3-nitro-6-chloropyridine and methane sulphonyl chloride inaccordance with the procedure described above for the preparation ofN-(6-chloro-3-nitro-2-pyridyl)acetamide.

HPLC: 8.08 min (90% pure)

MS: MH+=251.9 C6H6C1N3O4S 251 g/mol

2. Preparation of{6-[(2-{[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amino}ethyl)amino]-3-nitro(2-pyridyl)}(methylsulfonyl)amine

{6-[(2-{[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amino}ethyl)amino]-3-nitro(2-pyridyl)}(methylsulfonyl)aminewas prepared from[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-ylethylamine and(6-chloro-3-nitro(2-pyridyl))(methylsulfonyl)amine in accordance withthe procedure described above for the preparation of[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(6-methoxy-5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC: 10.8 min (85% pure)

MS: MH+=564 C21H19C12N9O4S 563 g/mol

Example 121 Preparation of{6-[(2-{[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl]amino}ethyl)amino]-3-nitro(2-pyridyl)}(methylsulfonyl)amine

{6-[(2-{[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl]amino}ethyl)amino]-3-nitro(2-pyridyl)}(methylsulfonyl)aminewas prepared from[4-(2,4-dichlorophenyl)-5-imidazol-1-ylpyrimidin-2-ylethylamine and(6-chloro-3-nitro(2-pyridyl))(methylsulfonyl)amine in accordance withthe procedure described above for the preparation of[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(6-methoxy-5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC: 11.1 min (85% pure)

MS: MH+=564 C21H19C12N9O4S 563 g/mol

Example 122 Preparation of(2-{6-[(2-{[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amino}ethyl)amino]-3-nitro(2-pyridyloxy)}ethyl)dimethylamine

1. Preparation of [2-(6-chloro-3-nitro(2-pyridyloxy))ethyl]dimethylamine

[2-(6-chloro-3-nitro(2-pyridyloxy))ethyl]dimethylamine was prepared from2,2-dimethylaminoethanol in accordance to the procedure described abovefor the preparation of 6-chloro-2-methoxy-3-nitro-pyridine.

HPLC: 4.9 min (65% pure)

MS: MH+=246.1 C9H12C1N3O3 245 g/mol

2. Preparation of(2-{6-[(2-{[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amino}ethyl)amino]-3-nitro(2-pyridyloxy)}ethyl)dimethylamine

(2-{6-[(2-{[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amino}ethyl)amino]-3-nitro(2-pyridyloxy)}ethyl)dimethylaminewas prepared from[4-(2,4-dichloro-phenyl)-5-imidazol-2-ylpyrimidin-2-ylethylamine and[2-(6-chloro-3-nitro(2-pyridyloxy))-ethyl]dimethylamine in accordancewith the procedure described above for the preparation of[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(6-methoxy-5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC: 8.5 min (85% pure)

MS: MH+=558.3 C24H25C12N9O3 557.1 g/mol

Example 123 Preparation of(2-{6-[(2-{[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl]amino}ethyl)amino]-3-nitro(2-pyridyloxy)}ethyl)dimethylamine

(2-{6-[(2-{[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl]amino}ethyl)amino]-3-nitro(2-pyridyloxy)}ethyl)dimethylaminewas prepared from[4-(2,4-dichlorophenyl)-5-imidazol-1-ylpyrimidin-2-ylethylamine and[2-(6-chloro-3-nitro(2-pyridyloxy))ethyl]dimethylamine in accordancewith the procedure described above for the preparation of[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(6-methoxy-5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC: 8.3 min (85% pure)

MS: MH+=558.3 C24H25C12N9O3 557.1 g/mol

Example 124 Preparation of (1Z)-1-aza-2-(6-chloro(3-pyridyl))-1-methoxyprop-1-ene

To a solution of 5-(2-chloropyidinyl)oxime methyl ether (540 mg, 3.2mmole) in acetonitrile(9 ml), ethylene diamine (2.8 ml, 42 mmole) wasadded. The reaction mixture was stirred at 85° C. overnight,concentrated under vacuo. The crude product was dissolved in aqueoussodium hydroxide (1 M, 15 ml) and extracted with ethyl acetate and amixture of acetonitrile/methanol (10:1). The organic extract was driedwith sodium sulfate to give(1Z)-1-aza-2-(6-chloro(3-pyridyl))-1-methoxyprop-1-ene as an yellow oil(547 mg, 75%).

HPLC: 1.8 min (75% pure)

MS: MH+=195.1 C9H14N4O 194.1 g/mol

Example 125 Preparation of(2-{[5-((1Z)-2-aza-2-methoxy-1-methylvinyl)(2-pyridyl)]amino}ethyl)[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl]amine

(2-{[5-((1Z)-2-aza-2-methoxy-1-methylvinyl)(2-pyridyl)]amino}ethyl)[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl]aminewas prepared from the guanidine (which was made from[2-(6-chloro-3-nitro(2-pyridyloxy))ethyl]dimethylamine and thecorresponding eneaminone).

HPLC: 9.3 min (85% pure)

MS: MH+=483.2 C22H20C12N8O 482 g/mol

Example 126 Preparation of{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl]amine

{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-imidazolyl-pyrimidin-2-yl]aminewas prepared from[4-(2,4-dichlorophenyl)-5-imidazol-1-ylpyrimidin-2-ylethylamine and2-chloro-5-nitro-6-aminopyridine in accordance with the proceduredescribed above for the preparation of[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(6-methoxy-5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC [Method AZ-S], 6.15 min (100%)

MS (m+H/z), 486.

Example 127 Preparation of6-[(2-{[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carbonitrile

6-[(2-{[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carbonitrilewas prepared from[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-ylethylamine and2-chloro-5-cyanopyridine in accordance with the procedure describedabove for the preparation of[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(6-methoxy-5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC [Method AZ-S], 5.94 min (100%);

MS (m+H/z), 451.

Example 128 Preparation of[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl]{2-[(5-methyl(7a-hydro-1,2,4-triazolo[1,5-a]pyrimidin-7-yl))amino]ethyl}amine

[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl]{2-[(5-methyl(7a-hydro-1,2,4-triazolo[1,5-a]pyrimidin-7-yl))amino]ethyl}aminewas prepared from[4-(2,4-dichlorophenyl)-5-imidazol-1-ylpyrimidin-2-ylethylamine and7-methyl-9-chloro-1,2,4-triazolo(1,5-a)pyrimidine in accordance with theprocedure described above for the preparation of[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(6-methoxy-5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC [Method AZ-S], 5.80 min (70%);

MS (m+H/z), 481.

Example 129 Preparation of[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl]{2-[(5-trifluoromethyl(2-pyridyl))amino]ethyl}amine

[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl]{2-[(5-trifluoromethyl(2-pyridyl))amino]ethyl}aminewas prepared from[4-(2,4-dichlorophenyl)-5-imidazol-1-ylpyrimidin-2-ylethylamine and2-chloro-5-trifluoromethylpyridine in accordance with the proceduredescribed above for the preparation of[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(6-methoxy-5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC. [Method AZ-S], 7.62 min (60%);

MS (m+H/z), 494.

Example 130 Preparation of4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl]{2-[(5-nitro(1,3-thiazol-2-yl))amino]ethyl}amine

4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl]{2-[(5-nitro(1,3-thiazol-2-yl))amino]ethyl}aminewas prepared from[4-(2,4-dichlorophenyl)-5-imidazol-1-ylpyrimidin-2-ylethylamine and2-chloro-5-nitro-thiazole in accordance with the procedure describedabove for the preparation of[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(6-methoxy-5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC [Method AZ-S], 7.14 min (100%);

MS (m+H/z), 477.

Example 131 Preparation of[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl]{2-[(6-chloropyrimidin-4-yl)amino]ethyl}amine

[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl]{2-[(6-chloropyrimidin-4-yl)amino]ethyl}aminewas prepared from[4-(2,4-dichlorophenyl)-5-imidazol-1-ylpyrimidin-2-ylethylamine and4,6-dichloropyrirnidine in accordance with the procedure described abovefor the preparation of[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(6-methoxy-5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC [Method AZ-S], 7.43 min (80%);

MS (m+H/z), 461.

Example 132 Preparation of[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl]{2-[(6-chlorobenzothiazo1-2-yl)amino]ethyl}amine

[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl]{2-[(6-chlorobenzothiazo1-2-yl)amino]ethyl}amine was prepared from[4-(2,4-dichlorophenyl)-5-imidazol-1-ylpyrimidin-2-ylethylamine and2,6-dichlorobenzothiazole in accordance with the procedure describedabove for the preparation of[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(6-methoxy-5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC [Method AZ-S], 8.23 min (100%);

MS (m+H/z), 516.

Example 133 Preparation of[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl]{2-[(3-nitro(2-thienyl))amino]ethyl}amine

[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl]{2-[(3-nitro(2-thienyl))amino]ethyl}aaminewas prepared from[4-(2,4-dichlorophenyl)-5-imidazol-1-ylpyrimidin-2-ylethylamine and2-chloro-3-nitrothiophene in accordance with the procedure describedabove for the preparation of[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(6-methoxy-5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC [Method AZ-S], 9.50 min (75%);

MS (m+H/z), 495.

Example 134 Preparation of4-amino-2-[(2-{[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl]amino}ethyl)amino]pyrimidine-5-carbonitrile

4-amino-2-[(2-{[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl]amino}ethyl)amino]pyrimidine-5-carbonitrilewas prepared from[4-(2,4-dichlorophenyl)-5-imidazol-1-ylpyrimidin-2-ylethylamine and2-chloro-4-amino-5-cyanopyrimidine in accordance with the proceduredescribed above for the preparation of[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(6-methoxy-5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC [Method AZ-S], 6.00 min (70%);

MS (m+H/z), 467.

Example 135 Preparation of{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[6-(2,4-dichlorophenyl)-5-nitro(2-pyridyl)]amine

1. Preparation of 2-(2,4-dichlorophenyl)-6-chloro-3-nitropyridine

1 mmol 2,6-dichloro-3-nitropyridine, 1.05 mmol of2,4-dichlorobenzeneboronic acid, and 3 mmol of Na₂CO₃, were dissolved in1.5 ml THF and 0.5 ml water and purged with nitrogen. 0.05 mmol of[1,1′-Bis(diphenylphosphino)ferrocene]dichloro-palladium(II) was addedto reaction and stirred at room temperature under nitrogen for fourteenhours. The reaction mixture was concentrated in vacuo and diluted withwater and ethyl acetate. The solution was extracted three times withethyl acetate, dried over sodium sulfate, and purified by columnchromatography eluting with 10% ethylacetate 90% hexanes to obtain2-(2,4-dichlorophenyl)-6-chloro-3-nitropyridine.

2. Preparation of (2-aminoethyl)(6-amino-5-nitro(2-pyridyl))amine

1 mmol of 2-amino-6-chloro-3-nitropyridine and 15 mmol of1,2-diaminoethane were stirred at reflux for fourteen hours. Thereaction mixture was concentrated in vacuo and solution of 1.5 mmol ofNaOH in water was added. This solution was extracted twice with 95%/5%methylene chloride/methanol. The aqueous was then saturated with saltand extracted twice with 95%/5% acetonitrile/methanol and then finallyextrated twice with 95%/5% ethylacetate/methanol. All organic fractionswere combined and dried over sodium sulfate to obtain(2-aminoethyl)(6-amino-5-nitro(2-pyridyl))amine.

3. Preparation of{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[6-(2,4-dichlorophenyl)-5-nitro(2-pyridyl)]amine

1 mmol of 2-(2,4-dichlorophenyl)-6-chloro-3-nitropyridine was taken with2 mmol of (2-aminoethyl)(6-amino-5-nitro(2-pyridyl))amine and 3 mmol ofN,N-diisopropylethylamine in 2 ml of DMF at 80° C. for two hours. Thereaction mixture was concentrated in vacuo and diluted with water andethyl acetate. The solution was extracted three times with ethylacetate, dried over sodium sulfate, and purified by columnchromatography eluting with 5-10% methanol/methylene chloride to obtain{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[6-(2,4-dichlorophenyl)-5-nitro(2-pyridyl)]amine.

HPLC: 8.698 min. (100% purity)

MS: MH+=464.1

Example 136 Preparation of6-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]-3-pyrrolino[3,4-b]pyridine-5,7-dione

1. Preparation of2-[2-(2,4-dichlorophenyl)-2-oxoethyl]isoindoline-1,3-dione

1 mmol of 2,4-dichlorophenacyl chloride in DMF was added drop wise to 2mmol of phthalimide and 2 mmol of Cs₂CO₃ in DMF at room temperature forfourteen hours and then purified by trituration with diethyl ether toobtain 2-[2-(2,4-dichlorophenyl)-2-oxoethyl]isoindoline-1,3-dione.

2. Preparation of2-{2-(2,4-dichlorophenyl)-1-[(dimethylamino)methylene]-2-oxoethyl}isoindoline-1,3-dione

1 mmol of 2-[2-(2,4-dichlcrophenyl)-2-oxoethyl]isoindoline-1,3-dion washeated to 80° C. in neat N,N-dimethylformamidedimethyl acetal for sixhours. The reaction mixture was concentrated in vacuo and purified bytrituration with diethyl ether to obtain2-{2-(2,4-dichlorophenyl)-1-[(dimethylamino)methylene]-2-oxoethyl}isoindoline-1,3-dione.

3. Preparation of2-{N-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]carbamoyl}benzoicacid

1 mmol of2-{2-(2,4-dichlorophenyl)-1-[(dimethylamino)methylene]-2-oxoethyl}isoindoline-1,3-dione,1 mmol of amino{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}carboxamidine, and 3 mmol ofCs₂CO₃ were dissolved in DMF and heated to 90° C. for fourteen hours.The reaction mixture was concentrated in vacuo and diluted with waterand ethyl acetate. The solution was extracted three times with ethylacetate and dried over sodium sulfate to obtain of2-{N-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]carbamoyl}benzoicacid.

4. Preparation of2-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]isoindoline-1,3-dione

1 mmol of2-{N-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]carbamoyl}benzoicacid was heated to 120° C. in acetic acid for four hours and thenconcentrated in vacuo to obtain2-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]isoindoline-1,3-dione.

5. Preparation of[5-amino-4-(2,4-dichlorophenyl)pyrimidin-2-yl]{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amine

1 mmol of2-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]isoindoline-1,3-dioneand 20 mmol of hydrazine were stirred in ethanol at 75° C. for two hoursand then purified by column chromatography eluting with 5-10%methanol/methylene chloride to obtain[5-amino-4-(2,4-dichlorophenyl)pyrimidin-2-yl]{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amine.

6. Preparation of6-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]-3-pyrrolino[3,4-b]pyridine-5,7-dione

1 mmol of[5-amino-4-(2,4-dichlorophenyl)pyrimidin-2-yl]{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amine,and 2 mmol of furano[3,4-b]pyridine-5,7-dione were stirred at roomtemperature for four hours. 2 mmol of HBTU, and 3 mmol ofN,N-diisopropylethylamine were added to solution and left for six hoursat room temperature. The reaction mixture was concentrated in vacuo anddiluted with water and ethyl acetate. The solution was extracted threetimes with ethyl acetate, dried over sodium sulfate, and purified bycolumn chromatography eluting with 5-10% methanol/methylene chloride toobtain6-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]-3-pyrrolino[3,4-b]pyridine-5,7-dione.

HPLC: 7.829 min. (97.32% purity)

MS: MH+=566.0

Example 137 Preparation of[6-(2,4-dichlorophenyl)-5-imidiazol-2-yl(2-pyridyl)]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine

1. Preparation of 1-(2,4-dichlorophenyl)-2-imidazol-2-ylethan-1-one.

The preparation of the material1-(2,4-dichlorophenyl)-2-imidazol-2-ylethan-1-one can be found in theprecedures for the preparation of[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine.

2. Preparation of 2-(2-aminoethylamino)-5-nitropyridine.

The preparation of the material 6-[(2-aminoethyl)amino]pyridine withvarious substitutents on the pyridine can be found in the precedures forthe preparation of[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine,Example 74, and its analogues.

3. Preparation of methyl 2-phenylthiopropanoate.

A solution of methyl-2-bromopropionate (6.13 M) in abs. ethanol (25 ml)was added slowly (˜1 h) to a stirred solution of thiophenol (107 M), KOH(107 M) in abs. ethanol (90 ml) at room temperature. After 12 h, thereaction was filtered and stripped of solvent under reduced pressure.The resulting slurry was partitioned with water (150 ml) and ether (100ml). The aqueous layer was extracted with ether (3×100 ml). The combinedorganic layers were washed with 1 M NaOH (2×50 ml), water (2×50 ml),brine (100 ml), dried with Mg₂SO₄, filtered, and concentrated underreduced pressure. Drying in vacuo for 2-3 hours results in methyl2-phenylthiopropanoate as a clear oil in 90% yield and >99% purity. (seeanalogous procedure in Warren, S.; et. al. J Chem. Soc. Perkin Trans. I1986, 1939-1945.)

4. Preparation of methyl 2-(phenylsulfinyl)propanoate.

A solution of mCPBA (57-86% active) in dry ether (200 ml, ˜0.25 g perml) was added dropwise to a stirred solution of methyl2-phenylthiopropanoate (0.34 M) in dry ether (400 ml) at 0° C. Thereaction was followed by TLC eluting with 10% ethyl acetate in hexane.After all of the starting material had been consumed, the reaction wasconcentrated under reduced pressure. The residue was dissolved in ether(150 ml) and dichloromethane (400 ml). The organics were washed with 1 MNa₂S₂O₃ (2×80 ml), sat. aq. Na₂CO₃ (4×100 ml), brine (100 ml), and driedwith Na₂SO₄. After removing the volatile organics, of methyl2-(phenylsulfinyl)propanoate was obtained in 99% yield and 99% purity.

5. Preparation of methyl 2-phenylthioprop-2-enoate.

A mixture of methyl 2-(phenylsulfinyl)propanoate (0.17 M) indichloromethane (800 ml), acetic anhydride (20 ml), and methanesulfonicacid (1.5 ml) were heated at 40° C. for 18 h. The reaction wasconcentrated under reduced pressure in a water bath that was held at aconstant 35° C. The residue was partitioned with water (200 ml) andether (100 ml). The aqueous layer was extracted with ether (3×50 ml).The combined organic layers were washed with water (50 ml), sat. aq.Na₂CO₃ (3×30 ml), brine (30 ml), and dried with MgSO₄. After filtration,the product was stripped of solvent under reduced pressure at <35° C.The product was purified by eluting through a short column of silica gelusing a 10% ethyl acetate in hexane mixture as eluent. Uponconcentration, methyl 2-phenylthioprop-2-enoate was isolated in 50%yield.

6. Preparation of methyl5-(2,4-dichlorophenyl)-4-imidazol-2-yl-5-oxo-2-phenylthiopentanoate.

A solution (1 M) of tert-butoxide in tert-butanol (54.9 ml) was added toa stirred solution of 1-(2,4-dichlorophenyl)-2-imidazol-2-ylethan-1-one(0.11 M) and methyl 2-phenylthioprop-2-enoate (0.14 M) dissolved indichloromethane (300 ml) and methanol (200 ml) at room temperature. Thereaction, which developed a dark color, was typically stirred overnight(ca. 16 h) under argon. The reaction was monitored by TLC using 5%methanol in dichloromethane as the solvent system. Additional methyl2-phenylthioprop-2-enoate was added as needed to consume all of thestarting 1-(2,4-dichlorophenyl)-2-imidazol-2-ylethan-1-one. The reactionwas quenched by the addition of sat. aq. NH₄Cl (˜100 ml). The mixturewas transferred to a separatory funnel and diluted with ethyl acetate(300 ml). The aqueous layer was removed and the organic layer was washedwith sat. aq. NH₄Cl (3×100 ml), brine (100 ml), and dried with Na₂SO₄.After filtration and evaporation, the residue was purified by flashchromatography over silica gel. The column was eluted starting with 100%dichloromethane to remove the non-polar methyl2-phenylthioprop-2-enoate. The product was eluted with 3% methanol indichloromethane. After drying overnight in vacuo, methyl5-(2,4-dichlorophenyl)-4-imidazol-2-yl-5-oxo-2-phenylthiopentanoate as adeep red glass was obtained in 71% yield. A further 10% yield of productcould be obtained by resubmitting side product contaminated fractions toa second purification by flash chromatography.

7. Preparation of6-(2,4-dichlorophenyl)-5-imidazol-2-yl-3-phenylthio-1,3,4-trihydropyridin-2-one.

A solution was made of methyl5-(2,4-dichlorophenyl)-4-imidazol-2-yl-5-oxo-2-phenylthiopentanoate(0.33 M), gla. acetic acid (21 ml), abs. ethanol (63 ml), toluene (21ml) by heating the mixture to 90° C. To the stirred solution, NH₄OAc(1.97 M) and 4 Å molecular sieves (15 g) were added. After 24 h ofheating, another portion of both NH₄OAc (1.97 M) and 4 Å molecularsieves (15 g) were added. After 48 h, the reaction had come tocompletion as determined by HPLC. The reaction is diluted with ethylacetate (500 ml), filtered, and washed with sat. aq. NaHCO₃ (4×250 ml)and brine (200 ml). The organic layer was dried with Na₂SO₄ andconcentrated under reduced pressure. The residue was redissolved inEtOAc and concentrated under reduced pressure. The residue was taken upin a minimal amount of EtOAc to precipitate the product. The remainingproduct could be purified by flash chromatography over silica gel. Thecolumn was eluted using 100% ethyl acetate. After concentration, theproduct was dried in vacuo, The product6-(2,4-dichlorophenyl)-5-imidazol-2-yl-3-phenylthio-1,3,4-trihydropyridin-2-oneobtained from both precipitation and chromatography gave a tan solid in89% yield.

8. Preparation of6-(2,4-dichlorophenyl)-5-imidazol-2-ylhydropyridin-2-one.

Dichloromethane (˜150 ml) was added carefully so as not to causeprecipitation to a solution of6-(2,4-dichlorophenyl)-5-imidazol-2-yl-3-phenylthio-1,3,4-trihydropyridin-2-one(0.17 M) in THF (40 ml). A solution of mCPBA (2.85 g, ˜16.5 mmol; 65-85%active) in dichloromethane (50 mg per 1 ml) is added dropwise to thestirred solution of phenylthiol above at −20° C. After addingapproximately 1 equivalent of oxidant, the reaction was allowed to warmto room temperature. Additional mCPBA is titrated into the reactionuntil all of the starting material is gone as judged by TLC eluting in5% MeOH in dichloromethane (R_(f) of the product is ˜0.1). As thereaction nears completion, the elimination product starts to precipitateout of solution as a gum. Upon completion, the reaction is stirred foran additional 30 min. Triethylamine (4 ml; 2 eq. based on mCPBA) isadded to the reaction which causes the reaction to go completely clearfor approximately 1 min. followed by nearly complete precipitation ofthe product as an off-white solid. The solid is filtered and washed withdichloromethane (3×30 ml). The product was dried in vacuo resulting inof 6-(2,4-dichlorophenyl)-5-imidazol-2-ylhydropyridin-2-one in 93%yield.

9. Preparation of6-(2,4-dichlorophenyl)-5-{1-[(trifluoromethyl)sulfonyl]imidazol-2-yl}-2-pyridyl(trifluoromethyl)sulfonate.

Trifluoromethanesulfonic anhydride (1.61 ml, 9.78 mmol) was added to astirred suspension of6-(2,4-dichlorophenyl)-5-imidazol-2-ylhydropyridin-2-one (500 mg, 1.63mmol) in pyridine (10 ml) at −10° C. After 30 min., the reaction wasallowed to warm to room temperature. Stirring continued until all of thesolid starting material had been dissolved and reacted as determined byHPLC. The reaction was diluted with dichloromethane (500 ml) and washedsequentially with sat. aq. NaHCO₃ (3×100 ml), water (2×100 ml), sat. aq.NaHCO₃ (100 ml), brine, dried with Na₂SO₄, filtered and concentratedunder reduced pressure. The residue was purified over a short column ofsilica gel eluting with 25% ethyl acetate in hexanes to remove thenon-polar product from baseline material. After removing the solventsand drying in vacuo, the product6-(2,4-dichlorophenyl)-5-{1-[(trifluoromethyl)sulfonyl]imidazol-2-yl}-2-pyridyl(trifluoro-methyl)sulfonate was obtained as a slightly yellow clearglass weighting 874 mg in 95% yield.

10. Preparation of[6-(2,4-dichlorophenyl)-5-imidazol-2-yl(2-pyridyl)]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine.

A suspension of (2-aminoethyl)(5-nitro(2-pyridyl))amine (255 mg, 1.40mmol) in N,N-dimethylacetamide (3 ml) containing dry powdered 4 Åmolecular sieves (50 mg) was added to a suspension of6-(2,4-dichlorophenyl)-5-{1-[(trifluoromethyl)sulfonyl]imidazol-2-yl}-2-pyridyl(trifluoromethyl)sulfonate (200 mg, 0.35 mmol) in N,N-dimethylacetamide(3 ml) containing dry powdered 4 Å molecular sieves (50 mg). Afterstirring at 40° C. for 24 h, ethylenediamine (0.5 ml) and water (0.5 ml)were added to the reaction to hydrolyze the one remaining triflate fromthe product. The reaction was stirred at 85° C. for 2 h and left at roomtemperature for 12 h. The reaction was then diluted with ethyl acetate(100 ml), filtered, extracted with sat. aq. NaHCO₃ (6×30 ml), brine (30ml), dried with Na₂SO₄, filtered, and concentrated under reducedpressure. The product[6-(2,4-dichlorophenyl)-5-imidazol-2-yl(2-pyridyl)]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine(79859) was obtained as a yellow glass weighing 143 mg in 85% yield.

HPLC: 22.1 min (>95% purity)

MS: M+H=470.2 (C₂₁H₁₇Cl₂N₇O₂+H=470)

Example 138 Preparation of give[4-(2,4-dichlorophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-5-yl]methan-1-ol

To a stirred solution of a suspension of 2.13 g (4.68 mmol) ethyl4-(2,4-dichloro-phenyl)-2-({2-[(tert-butoxy)carbonylamino]ethyl}amino)pyrimidine-5-carboxylatein 10 mL THF at room temperature under nitrogen was added 25 mL ofDIBAL-H (1 M in THF, 25.0 mmol) dropwise. During this addition thesuspension gradually turned into a homogenous yellow solution. After 1 hthe resulting solution was heated to 70° C. for an additional 7 h. Thereaction was then cooled and the reaction was quenched by the additionof Rochelle's salt. The resulting suspension was partitioned betweenmethylene chloride and water. The aqueous layer was extracted twice withmethylene chloride and the combined organic layers washed with brine anddried with sodium sulfate. Concentration gave 2.05 g of a yellow foam.Chromatography on silica gel (110 g) using 5% methanol/ether as eluentgave 430 mg (22%) ofN-(2-{[4-(2,4-dichlorophenyl)-5-(hydroxymethyl)pyrimidin-2-yl]amino}ethyl)(tert-butoxy)carboxamideas a colorless foam.

HPLC [Method AZ-S], 9.42 min (100%); MS (m+H/z), 413.

N-(2-{[4-(2,4-dichlorophenyl)-5-(hydroxymethyl)pyrimidin-2-yl]amino}ethyl)(tert-butoxy)carboxamide(372 mg, 0.90 mmol) was dissolved in 2 mL of anhdyrous trifluoroaceticacid and stirred at room temperature for 2 h. Evaporation of the solventafforded{2-[(2-aminoethyl)amino]-4-(2,4-dichlorophenyl)pyrimidin-5-yl}methan-1-ol,as its trifluoracetate salt, in quantitative yield.

{2-[(2-aminoethyl)amino]-4-(2,4-dichlorophenyl)pyrimidin-5-yl}methan-1-olwas dissolved in 3 mL of anhydrous THF and 1.47 g (4.50 mmol) ofanhydrous cesium carbonate was added. 2-chloro-5-nitro-6-aminopyridine(143 mg, 0.9 mmol) was added in one portion and the yellow suspensionheated at 70° C. for 18 h. The reaction mixture was filtered,concentrated and the residue chromatographed (silica gel, 5%methanol/methylene chloride) to give[4-(2,4-dichlorophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-5-yl]methan-1-ol,216 mg (53%), as a yellow solid.

HPLC [Method AZ-S], 6.92 min (100%); MS (m+H/z), 450.

Example 139 Preparation of[4-(2,4-dichlorophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-5-yl]methan-1-ol

The process of Example 140 is repeated using 2-chloro-5-nitro-pyridineand{2-[(2-aminoethyl)amino]-4-(2,4-dichlorophenyl)pyrimidin-5-yl}methan-1-ol.Chromatography of the residue (silica gel, 5% methanol/methylenechloride) afforded 200 mg (51%) of[4-(2,4-dichlorophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-5-yl]methan-1-olas a yellow solid.

HPLC [Method AZ-S], 7.85 min (100%); MS (m+H/z), 435.

Example 140 Preparation of[4-(2,4-dichlorophenyl)-5-(morpholin-4-ylmethyl)pyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine

Using the method of Swern, et al., 100 mg ofN-(2-{[4-(2,4-dichlorophenyl)-5-(hydroxymethyl)pyrimidin-2-yl]amino}ethyl)(tert-butoxy)carboxamidewas dissolved in 1 mL of anhydrous methylene chloride and added to asolution of oxalyl chloride (30.7 μL, 0.363 mmol) and DMSO (51.6 μL,0.726 mmol) which had been stirring at −78° C. for 15 min. The resultingsolution was stirred for an additional 30 min, at which time 202 μL(1.45 mmol) of triethyl amine was added. The resulting suspension wasallowed to warm to room temperature after 15 min and 1 mL of water wasadded and the layers separated. The aqueous layer was extracted withmethylene chloride and the combined organic layers dried (sodiumsulfate) and concentrated to affordN-(2-{[4-(2,4-dichlorophenyl)-5-formylpyrimidin-2-yl]amino}ethyl)(tert-butoxy)carboxamideas a light yellow foam. This product proved to be air sensitive and wasused without further manipulation.

HPLC [Method AZ-S], 11.41 min (95%); MS (m+H/z), 411.

N-(2-{[4-(2,4-dichlorophenyl)-5-formylpyrimidin-2-yl]amino}ethyl)(tert-butoxy)carboxamide(50 mg, 0.121 mmol) was dissolved in 5 mL of THF, 242 μL of sodiumcyanoborohydride (1 M in THF) and 5 μL of glacial acetic acid added andthe mixture was heated to 70° C. for 18 h. Following slow addition of 1mL of water to decompose excess reagent, the mixture was partitionedbetween ethyl acetate and saturated citric acid solution. The organiclayer was discarded, and aqueous layer was carefully basified withsodium hydoxide to PH 9, then extracted with ethyl acetate twice. Thecombined organic layers was dried (sodium sulfate), concentrated, andpurified by chromatography (selica gel, 10% methanol/methylene chloride)to affordN-(2-{[4-(2,4-dichlorophenyl)-5-(morpholin-4-ylmethyl)pyrimidin-2-yl]amino}ethyl)(tert-butoxy)carboxamide,30 mg (52%).

HPLC [Method AZ-S], 8.28 min (95%); MS (m+H/z), 482.

Using the conditions described in Step 1.2 above,N-(2-{[4-(2,4-dichlorophenyl)5-(morpholin-4-ylmethyl)pyrimidin-2-yl]amino}ethyl)(tert-butoxy)carboxamidewas treated with anhydrous trifluouroacetic acid to afford(2-aminoethyl)[4-(2,4-dichlorophenyl)-5-(morpholin-4-ylmethyl)pyrimidin-2-yl]aminein near quantitative yield.

HPLC [Method AZ-S], 3.97 min (95%); MS (m+H/z), 382.

Using the conditions described in the previous example,(2-aminoethyl)[4-(2,4-dichlorophenyl)-5-(morpholin-4-ylmethyl)pyrimidin-2-yl]amineand 9.7 mg (0.061 mmol) of 2-chloro-5-nitropyridine gave[4-(2,4-dichlorophenyl)-5-(morpholin-4-ylmethyl)pyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}aminein 60% yield following chromatography (silica gel, 5% methanol/methylenechloride).

HPLC [Method AZ-S], 7.43 min (100%); MS (m+H/z), 504.

Example 141 Preparation of[4-(2,4-dichlorophenyl)-5-(morpholin-4-ylmethyl)pyrimidin-2-yl]{2-[(5-nitro-6-amino(2-pyridyl))amino]ethyl}amine

Using the conditions described in Example 139,(2-aminoethyl)[4-(2,4-dichlorophenyl)-5-(morpholin-4-ylmethyl)pyrimidin-2-yl]amineand 10.6 mg (0.061 mmol) of 2-chloro-5-nitro-6-aminopyridine afforded191 mg (60%) of[4-(2,4-dichlorophenyl)-5-(morpholin-4-ylmethyl)pyrimidin-2-yl]{2-[(5-nitro-6-amino(2-pyridyl))amino]ethyl}amineas a yellow solid following chromatography (silica gel, 5%methanol/methylene chloride).

HPLC [Method AZ-S], 6.49 min (100%); MS (m+H/z), 519.

Example 142 Preparation of[4-(2,4-dichlorophenyl)-5-(morpholin-4-ylmethyl)pyrimidin-2-yl]{2-[(5-nitro-6-amino(2-pyridyl))amino]ethyl}amineand related compounds

A suspension of 0.44 g (2.59 mmol) of2,2,2-trifluoro-N-[2-(methylamino)ethyl]acetamide (prepared according toSyn. Comm., 26:3633-3636 (1996)) and 0.38 g (2.59 mmol) of1-pyrazolocarboxamidine hydrochloride in 5 mL anhydrous THF was stirredat room temperature for 3 h. Concentration of this suspension afforded awhite solid which was found by 1H NMR analysis to consist of the desiredguanidine, N-[2-(amidinomethylamino)ethyl]-2,2,2-trifluoroacetamidehydrochloride, and pyrazole. This was used without further purificationin the next step.

A solution of 0.60 g (2.3 mmol) 2,4-dichloro-2-(1-imidazoyl)ethan-1-one,0.38 mL (2.82 mmol) dimethyformamide dimethylacetal and 5 mL THF wasrefluxed for 2 h. Concentration afforded1-(2,4-dichlorophenyl)-2-(1-imidazoyl)-3-dimethylaminoprop-2-en-1-one asa light red solid in quantitative yield. This solid was redissolved in 5mL THF, 1.0 g (3.06 mmol) of anhydrous cesium carbonate and the residuecontaining N-[2-(amidinomethylamino)ethyl]-2,2,2-trifluoroacetamidehydrochloride described above were added and the resulting mixtureheated to 70° C. for 18 h. After cooling, water was added and theresulting mixture extracted with methylene chloride. The aqueous layerwas extracted with methylene chloride and the combined organics washedwith brine, dried and concentrated to afford 1.56 g of a brown oil.Chromatography (silica gel, 5% methanol/methylene chloride) afforded0.35 g of the desired pyrimidine,N-(2-{[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl]methylamino}ethyl)-2,2,2-trifluoroacetamideas a brown solid.

HPLC [Method AZ-S], 7.68 min (85%); MS (m+H/z), 459.

The aforementioned pyrimidine,N-(2-{[4-(2,4-dichlorophenyl)-5-imidazolyl-pyrimidin-2-yl]methylamino}ethyl)-2,2,2-trifluoroacetamide(114 mg, 0.25 mmol) was dissolved in 2 mL methanol and potassiumhydroxide (40 mg, 1 mmol) added. This suspension was stirred at roomtemperature for 1 hour. Water was added, and the solution extracted withmethylene chloride. The aqueous layer was thoroughly extracted withmethylene chloride and the organic layers washed with brine, dried andconcentrated to give the deprotected primary amine,(2-aminoethyl)[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl]methylamine,in quantitative yield.

HPLC [Method AZ-S], 4.43 min (90%); MS (m+H/z), 363.

Using the procedure described above in Example 139, reaction of(2-aminoethyl)[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl]methylamine and2-chloro-5-nitropyridine were reacted to afford[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl]methyl{2-[(5-nitro(2-pyridyl))amino]ethyl}amine(73174).

HPLC [Method AZ-S], 7.82 min (100%); MS (m+H/z), 485.

Using the procedure described above, reaction of(2-aminoethyl)[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl]methylamineand 2-chloro-5-nitro-6-amino-pyridine were reacted to afford[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl]methyl{2-[(5-nitro-6-amino(2-pyridyl))amino]ethyl}amine.

HPLC [Method AZ-S], 6.73 min (100%); MS (m+H/z), 500.

Using the procedure described above in Example 139, reaction of(2-aminoethyl)[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl]methylamine and2-chloro-5-cyanopyridine were reacted to afford[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl]methyl{2-[(5-cyano(2-pyridyl))amino]ethyl}amine.

HPLC [Method AZ-S], 6.49 min (100%); MS (m+H/z), 465.

A solution of 0.60 g (2.3 mmol) 2,4-dichloro-2-(2-imidazoyl)ethan-1-one,0.38 mL (2.82 mmol) dimethyformamide dimethylacetal and 5 mL THF wasrefluxed for 2 h. Concentration afforded1-(2,4-dichlorophenyl)-2-(2-imidazoyl)-3-(dimethylamino)prop-2-en-1-oneas a light red solid in quantitative yield. This solid was redissolvedin 5 mL THF, 1.0 g (3.06 mmol) of anhydrous cesium carbonate andN-[2-(amidinomethylamino) ethyl]-2,2,2-trifluoroacetamide hydrochloride(vide supra) were added and the resulting mixture heated to 70° C. for18 h. After cooling, water was added and the resulting mixture extractedwith methylene chloride. The aqueous layer was extracted with methylenechloride and the combined organics washed with brine, dried andconcentrated to afford a brown oil. Chromatography (silica gel, 5%methanol/methylene chloride) afforded 0.30 g of the desired pyrimidine,N-(2-{[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]methylamino}ethyl)-2,2,2-trifluoroacetamide,as a brown solid.

HPLC [Method AZ-S], 7.25 min (100%); MS (m+H/z), 459.

The aforementioned pyrimidine,N-(2-{[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]methylamino}ethyl)-2,2,2-trifluoroacetamide(114 mg, 0.25 mmol) was dissolved in 2 mL methanol and potassiumhydroxide (40 mg, 1 =mol) added. This suspension was stirred at roomtemperature for 1 hour. Water was added, and the solution extracted withmethylene chloride. The aqueous layer was thoroughly extracted withmethylene chloride and the organic layers washed with brine, dried andconcentrated to give the deprotected primary amine,(2-aminoethyl)[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]methylaminein quantitative yield.

HPLC [Method AZ-S], 3.92 min (100%); MS (m+H/z), 363.

Using the procedure described above in Example 139, reaction of(2-aminoethyl)[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]methylamineand 2-chloro-5-cyano-pyridine were reacted to afford[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]methyl{2-[(5-cyano(2-pyridyl))amino]ethyl}amine.

HPLC [Method AZ-S], 5.98 min (100%); MS (m+H/z), 465.

Using the procedure described above in Example 139, reaction of(2-aminoethyl)[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]methylamineand 2-chloro-5-nitro-pyridine were reacted to afford[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]methyl{2-[(5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC [Method AZ-S], 7.31 min (100%); MS (m+H/z), 485.

Using the procedure described above in Example 139, reaction of(2-aminoethyl)[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]methylamineand 2-chloro-5-nitro-6-aminopyridine were reacted to afford[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]methyl{2-[(5-nitro-6-amino(2-pyridyl))amino]ethyl}amine.

HPLC [Method AZ-S], 5.85 min (100%); MS (m+H/z), 500.

Example 143 Preparation of{2-[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yloxy]ethyl}(5-nitro(2-pyridyl))amine(76062) and related compounds

A suspension of 1.50 g (4.84 mmol)1-(2,4-dichlorophenyl)-2-(1-imidazoyl)-3-dimethylaminoprop-2-en-1-one(prepared as described in Step 3.1 above), 0.673 g (2.42 mmol) ofS-methylisothiourea nitrate and 2.05 g (6.29 mmol) in 30 mL ofN-methylpyrrolidinone (NMP) was heated to 80° C. for 2 h. Water wasadded and the mixture extracted with ethyl acetate. The aqueous layerwas further extracted with additional ethyl acetate. The combinedorganic layers were thoroughly washed with water, brine and dried withsodium sulfate. Concentration and chromatography of the residue (silicagel, 2% methanol/methylene chloride) afforded 800 mg (50%) of thedesired pyrimidine,4-(2,4-dichlorophenyl)-5-imidazolyl-2-methylthiopyrimidine.

HPLC [Method AZ-S], 7.40 min (100%); MS (m+H/z), 337.

4-(2,4-dichlorophenyl)-5-imidazolyl-2-methylthiopyrimidine (219 mg, 0.65mmol) was dissolved in 2 mL anhydrous methylene chloride and 590 mg(57-86%, 1.95 mmol) of m-chloroperoxybenzoic acid added at roomtemperature and stirred for 2 h. Saturated sodium carbonate was addedand the organic layer separated and washed with 10% aqueous sodiumsulfite. Drying (sodium sulfate) and concentration afforded 240 mg(100%) of4-(2,4-dichlorophenyl)-5-imidazolyl-2-(methylsulfonyl)pyrimidine as ayellow solid which was used without further purification.

HPLC [Method AZ-S], 5.47 min (100%); MS (m+H/z), 369.

To a stirred suspension of 1.58 g 2-chloro-5-nitropyridine (10.0 mmol)in 10 mL acetonitrile was added 1.81 mL (30 mmol) of ethanolaminedropwise at room temperature. After heating at 80° C. for 0.5 h, thereaction was cooled, water was added, followed by the addition of ether.Cooling this biphasic mixture at 5° C. led to the formation of a yellowsolid, which was collected and identified as2-[(5-nitro-2-pyridyl)amino]ethan-1-ol.

HPLC [Method AZ-S], 1.74 min (100%); MS (m+H/z), 184.

1.74 g 2-chloro-5-nitro-6-aminopyridine (10.0 mmol) was reacted at 80°C. with 1.81 mL (30 mmol) of ethanolamine. Cooling the reaction mixtureled to the formation of a yellow solid, which was collected andidentified as 2-[(6-amino-5-nitro-2-pyridyl)amino]ethan-1-ol.

HPLC [Method AZ-S], 1.32 min (100%); MS (m+H/z), 199.

1.38 g 2-chloro-5-cyanopyridine (10.0 mmol) was reacted with 1.81 mL (30mmol) of ethanolamine at 80° C. for 0.5 h. The reaction was cooled,water was added, followed by the addition of ether. Cooling thisbiphasic mixture at 5° C. led to the formation of a yellow solid, whichwas collected and identified as6-[(2-hydroxyethyl)amino]pyridine-3-carbonitrile.

HPLC [Method AZ-S], 1.13 min (100%); MS (m+H/z), 164.

To a stirred solution of 37.2 mg (0.203 mmol) of2-(5-nitro-2-aminopyridyl)ethanolamine in 1 mL anydrous THF at roomtemperature was added 244 μL of a 1M solution of sodiumhexamethyldisilazide (1M in toluene, 0.244 mmol). This solution wasstirred for 1 h, and a solution of4-(2,4-dichlorophenyl)-5-imidazolyl-2-(methylsulfonyl)pyrimidine in 1 mLanhydrous THF was added dropwise. After stirring for 4 h, water wasadded and the reaction mixture thoroughly extracted with ethyl acetate.The combined organics were washed with brine, dried (sodium sulfate),concentrated and chromatographed (silica gel, 5% methanol/methylenechloride) to give 15.7 mg of{2-[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yloxy]ethyl}(5-nitro(2-pyridyl))amine(76062) as a yellow solid.

HPLC [Method AZ-S], 7.44 min (85%); MS (m+H/z), 472.

As described above,{2-[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yloxy]ethyl}(5-nitro-6-amino(2-pyridyl))amine(76063) was synthesized using4-(2,4-dichlorophenyl)-5-imidazolyl-2-(methylsulfonyl)pyrimidine and2-(5-nitro-6-amino-2-pyridyl)ethanolamine to afford 24.3 mg of{2-[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yloxy]ethyl}(5-nitro-6-amino(2-pyridyl))amineas a light yellow solid.

HPLC [Method AZ-S], 6.47 min (90%); MS (m+H/z), 487.

As described in above,{2-[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yloxy]ethyl}(5-cyano(2-pyridyl))amine(76064) was synthesized using4-(2,4-dichloro-phenyl)-5-imidazolyl-2-(methylsulfonyl)pyrimidine and6-[(2-hydroxyethyl)amino]pyridine-3-carbonitrile to afford 27.6 mg of{2-[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yloxy]ethyl}(5-cyano(2-pyridyl))amineas a light yellow solid.

HPLC [Method AZ-S], 6.37 min (95%); MS (m+H/z), 452.

Example 144 Preparation of[2-(dimethylamino)ethoxy]-N-[4-(4-cyanophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-5-yl]carboxamide

A solution of 50 mg (0.12 mmol) of4-(4-cyanophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylicacid, 26.2 ide (0.12 mmol, DPPA), 17.2 μL of triethylamine (0.12 mmol)in 2 mL of THF was heated to 75° C. for 24 hours. After cooling, thesolution was concentrated and the residue chromatographed (silica, 5%methanol/methylene chloride) to give 40.2 mg (68%) of the desiredcarbamate,[2-(dimethylamino)ethoxy]-N-[4-(4-cyanophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-5-yl]carboxamide,as a colorless solid.

HPLC [Method AZ-S], 6.23 min (100%); MS (m+H/z), 492.

Example 145 Preparation of Additional Compounds

The compounds described in detail below were synthesized using thefollowing general procedures:

Step A. Alkylation

1 mmol of aryl substituted phenacyl chloride in DMF was added drop wiseto 2 mmol of amine and 2 mmol of Cs₂CO₃ in DMF at room temperature forfourteen hours. The reaction mixture was concentrated in vacuo anddiluted with water and ethyl acetate. The solution was extracted threetimes with ethyl acetate, dried over sodium sulfate, and purified bycolumn chromatography or trituration.

Step B. Enaminone Formation

1 mmol of substrate was heated to 80° C. in neat DMF-DMA for six hours.Product was concentrated in vacou and purified by trituration withdiethyl ether.

Step C. Pyrimidine Formation

1 mmol of substrate, 1 mmol of guanadine, and 3 mmol of Cs₂CO₃ wasdissolved in DMF and heated to 90° C. for fourteen hours. The reactionmixture was concentrated in vacuo and diluted with water and ethylacetate. The solution was extracted three times with ethyl acetate,dried over sodium sulfate, and purified by column chromatography with5-10% methanol in methylenechloride.

Step D. Cyclization to Imide

1 mmol of substrate was heated to 120° C. in acetic acid for four hoursand then concentrated in vacuo and purified by column chromatographywith 5-10% methanol in methylenechloride.

Step E. Phthalimide Cleavage

1 mmol of substrate and 20 mmol of hydrazine were stirred in ethanol at75° C. The Ethanol was removed from the reaction mixture in vacuo andthen methylene chloride was added the solution was filtered. Thefiltrate was collected and concentrated in vacuo.

Step F. Acid Coupling

1 mmol of substrate, 2 mmol of carboxylic acid, 2 mmol of HBTU, and 3mmol of Diisopropylethyl amine were stirred in THF. The reaction mixturewas concentrated in vacuo and diluted with water and ethyl acetate. Thesolution was extracted three times with ethyl acetate, dried over sodiumsulfate, and purified by column chromatography with 5-10% methanol inmethylenechloride.

Step G. Boc Deprotection

1 mmol of substrate was stir in a mixture of 1 ml methylenechloride and1 ml trifloroacetic acid at 40° C. for 30 min. and concentrated invacuo.

Step H. SnAr Tail Piece

1 mmol of substrate, 1.5 mmol of a substituted 2-chloropyridine, and 4mmol of Diisopropylethyl amine were stirred in 2 ml of DMF at 80° C. forfourteen hours. The reaction mixture was concentrated in vacuo anddiluted with water and ethyl acetate. The solution was extracted threetimes with ethyl acetate, dried over sodium sulfate, and purified bycolumn chromatography with 5-10% methanol in methylenechloride.

Step I. Bromination

20 mmol of aryl substituted acetiphenone, 1 ml conc. HCl were mixed in20 ml diethyl ether at 0° C. under nitrogen. To this solution a solutionof 20 mmol Br₂ in 20 ml chloroform was added drop wise and left for fourhours and then concentrated in vacuo.

Step J. SnAr on Ketone

1 mmol of 1-(4-fluorophenyl)-2-imidazolylethan-1-one, 0.3 mmol of anamine, and 1 mmol of K₂CO₃ were heated at 100° C. for 14 hours. Thereaction mixture was poured over ice, then filtered, and the solid wascollected.

Step K. Anhydride Coupling

1 mmol of substrate and 1 mmol of anhydride were stirred at roomtemperature for four hours in THF. The reaction mixture was concentratedin vacuo and diluted with water and ethyl acetate. The solution wasextracted three times with ethyl acetate, dried over sodium sulfate, andpurified by column chromatography eluting with 5-10% methanol/methylenechloride.

Step L. Suzuki Reaction

1 mmol of a 2,6-dichloro-pyridine, 1.05 mmol of boronic acid, and 3 mmolof Na₂CO₃ were dissolved in 1.5 ml THF and 0.5 ml water and purged withnitrogen. 0.05 mmol of[1,1′-Bis(diphenylphosphino)ferrocne]dichloropalladium(II) was added toreaction and stirred at room temperature under nitrogen for 14 hours.The reaction mixture was diluted with water and ethyl acetate. Thesolution was extracted three times with ethyl acetate, dried over sodiumsulfate, and purified by column chromatography with 10% ethylacetate 90%hexanes.

Step M. SnAr Reaction

1 mmol of substrate is taken with 2 mmol of amine and 3 mmol ofDiisopropylethyl amine in 2 ml of DMF at 80° C. for two hours. Thereaction mixture was concentrated in vacuo and diluted with water andethyl acetate. The solution was extracted three times with ethylacetate, dried over sodium sulfate, and purified by columnchromatography 30% ethylacetate 70% hexanes.

Step N. Nitro Reduction

1 mmol of substrate was taken with an equal weight of 5% Pd-C with 20mmol N₂H₄ and dissolved in THF. The reaction was stirred at reflux for24 hrs and then filtered through celite and purified by columnchromatography.

Step O. Ethanol Nitro Reduction

1 mmol of substrate was taken with an equal weight of 5% Pd-C anddissolved in ethanol. The reaction was placed in a Parr shaker under 35PSI of hydrogen for six hours, then filtered through celite, andpurified by column chromatography.

Example 145-4 Preparation of[4-(2,4-difluorophenyl)-5-imidazolylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine

[4-(2,4-difluorophenyl)-5-imidazolylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}aminewas made in accordance with the foregoing procedures using1-(2,4-difluorophenyl)-2-chloroethan-1-one and imidazole in step A, stepB and amino{2-[(5-nitro(2-pyridyl))amino]ethyl}carboxamidine in step C.

HPLC: 7.150 min.

MS: MH⁺=439.1

Example 145-2 Preparation of[5-imidazolyl-4-(4-methylphenyl)pyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine

[5-imidazolyl-4-(4-methylphenyl)pyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}aminewas made in accordance with the foregoing procedures using2-bromo-1-(4-methylphenyl)ethan-1-one and imidazole in step A, step Band amino{2-[(5-nitro(2-pyridyl))amino]ethyl}carboxamidine in step C.

HPLC: 7.333 min.

MS: MH⁺=417.2

Example 145-3 Preparation of[4-(2-chlorophenyl)-5-imidazolylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine

[4-(2-chlorophenyl)-5-imidazolylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}aminewas made in accordance with the foregoing procedures through step I,using 1-(2-chlorophenyl)ethan-1-one and imidazole in step A, step B andamino{2-[(5-nitro(2-pyridyl))amino]ethyl}carboxamidine in step C.

HPLC: 7.233 min.

MS: MH³⁰ =437.1

Example 145-4 Preparation of tert-butyl4-{4-[5-imidazolyl-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}piperazinecarboxylate

tert-butyl4-{4-[5-imidazolyl-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}piperazinecarboxylatewas made in accordance with the foregoing procedures using2-bromo-1-(4-fluorophenyl)ethan-1-one and imidazole in step A, step Jusing tert-butyl piperazinecarboxylate, step B andamino{2-[(5-nitro(2-pyridyl))amino]ethyl}carboxamidine in step C.

HPLC: 9.670 min.

MS: MH⁺=587.2

Example 145-5 Preparation of{5-imidazolyl-4-[4-(trifluoromethyl)phenyl]pyrimidin-2-yl}{2-[(5-nitro(2-pyridyl))amino]ethyl}amine

{5-imidazolyl-4-[4-(trifluoromethyl)phenyl]pyrimidin-2-yl}{2-[(5-nitro(2-pyridyl))amino]ethyl}aminewas made in accordance with the foregoing procedures through step Iusing 1-[4-(trifluoromethyl)phenyl]ethan-1-one, A using imidazole, B,and C using amino{2-[(5-nitro(2-pyridyl))amino]ethyl}carboxamidine.

HPLC: 8.533 min.

MS: MH⁺=471.2

Example 145-6 Preparation of[4-(4-ethylphenyl)-5-imidazolylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine

[4-(4-ethylphenyl)-5-imidazolylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}aminewas made in accordance with the foregoing procedures through steps Iusing 1-(4-ethylphenyl)ethan-1-one, A using imidazole, B, and C usingamino{2-[(5-nitro(2-pyridyl))amino]ethyl}carboxamidine.

HPLC: 8.267 min.

MS: MH⁺=431.2

Example 145-7 Preparation of[4-(3,5-dichloro(2-thienyl))-5-imidazolylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine

[4-(3,5-dichloro(2-thienyl))-5-imidazolylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}aminewas made in accordance with the foregoing procedures through steps Iusing 1-(3,5-dichloro-2-thienyl)ethan-1-one, A using imidazole, B, and Cusing amino{2-[(5-nitro(2-pyridyl))amino]ethyl}carboxamidine.

HPLC: 8.167 min.

MS: MH⁺=477.1

Example 145-8 Preparation of[5-imidazolyl-4-(4-piperazinylphenyl)pyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine

[5-imidazolyl-4-(4-piperazinylphenyl)pyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}aminewas made in accordance with the foregoing procedures through steps Ausing 2-bromo-1-(4-fluorophenyl)ethan-1-one and imidazole, J usingtert-butyl piperazinecarboxylate, B, C usingamino{2-[(5-nitro(2-pyridyl))amino]ethyl}carboxamidine, and G.

HPLC:

MS: MH⁺=487.3

Example 145-9 Preparation of2-{N-[4-(2,4-dichlorophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-5-yl]carbamoyl}benzoicacid

2-{N-[4-(2,4-dichlorophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-5-yl]carbamoyl}benzoicacid was made in accordance with the foregoing procedures through stepsA using 1-(2,4-dichlorophenyl)-2-chloroethan-1-one and phthalimide, B,and C using amino{2-[(5-nitro(2-pyridyl))amino]ethyl}carboxamidine.

HPLC: 11.433 min.

MS: MH⁺=568.1

Example 145-10 Preparation of5-[5-imidazolyl-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]thiophene-2-cabonitrile

5-[5-imidazolyl-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]thiophene-2-carbonitrilewas made in accordance with the foregoing procedures through steps Iusing 5-acetylthiophene-2-carbonitrile, A using imidazole, B, and Cusing amino {2-[(5-nitro(2-pyridyl))amino]ethyl}carboxamidine.

HPLC 7.517 min.

MS: MH⁺=434.1

Example 145-11 Preparation of{5-imidazolyl-4-[4-(4-methylpiperazinyl)phenyl]pyrimidin-2-yl}{2-[(5-nitro(2-pyridyl))amino]ethyl}amine

{5-imidazolyl-4-[4-(4-methylpiperazinyl)phenyl]pyrimidin-2-yl}{2-[(5-nitro(2-pyridyl))amino]ethyl}aminewas made in accordance with the foregoing procedures through steps Ausing 2-bromo-1-(4-fluorophenyl)ethan-1-one and imidazole, J usingmethylpiperazine, B, and C usingamino{2-[(5-nitro(2-pyridyl))amino]ethyl}carboxamidine.

HPLC: 5.417 min.

MS: MH⁺=501.3

Example 145-12 Preparation of[5-imidazolyl-4-(4-piperidylphenyl)pyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine

[5-imidazolyl-4-(4-piperidylphenyl)pyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}aminewas made in accordance with the foregoing procedures through steps Ausing 2-bromo-1-(4-fluorophenyl)ethan-1-one and imidazole, J usingpiperidine, B, and C usingamino{2-[(5-nitro(2-pyridyl))amino]ethyl}carboxamidine.

HPLC: 6.300 min.

MS: MH⁺=486.2

Example 145-13 Preparation ofN-{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}-N-[4-(2,4-dichlorophenyl)-5-(3,5-dioxomorpholin-4-yl)pyrimidin-2-yl]acetamide

N-{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}-N-[4-(2,4-dichlorophenyl)-5-(3,5-dioxomorpholin-4-yl)pyrimidin-2-yl]acetamidewas made in accordance with the foregoing procedures through steps Ausing 1-(2,4-dichlorophenyl)-2-chloroethan-1-one and phthalimide, B, Cusing amino{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}carboxamidine, D,E, and K using 1,4-dioxane-2,6-dione and then an excess of aceticanhydride.

HPLC: 13.117 min.

MS: MH⁺=575.1

Example 145-14 Preparation ofN-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]acetamide

N-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]acetamidewas made in accordance with the foregoing procedures through steps Ausing 1-(2,4-dichlorophenyl)-2-chloroethan-1-one and phthalimide, B, Cusing amino{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}carboxamidine, D,E, and K using acetic anhydride.

HPLC: 10.200 min.

MS: MH⁺=477.0

Example 145-15 Preparation of4-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]morpholine-3,5-dione

4-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]morpholine-3,5-dionewas made in accordance with the foregoing procedures through steps stepsA using 1-(2,4-dichlorophenyl)-2-chloroethan-1-one and phthalimide, B, Cusing amino{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}carboxamidine, D,E, K using 1,4-dioxane-2,6-dione, and F.

HPLC: 9.317 min.

MS: MH⁺=533.1

Example 145-16 Preparation ofN-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]-2-(dimethylamino)acetamide

N-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]-2-(dimethylamino)acetamidewas made in accordance with the foregoing procedures through steps Ausing 1-(2,4-dichlorophenyl)-2-chloroethan-1-one and phthalimide, B, Cusing amino{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}carboxamidine, D,E, and F using 2-(dimethylamino)acetic acid.

HPLC: 6.567 min.

MS: MH⁺=520.2

Example 145-17 Preparation of1-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]-3-pyrroline-2,5-dione

1-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]-3-pyrroline-2,5-dionewas made in accordance with the foregoing procedures through steps Ausing 1-(2,4-dichlorophenyl)-2-chloroethan-1-one and phthalimide, B, Cusing amino{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}carboxamidine, D,E, K using malic anhydride, and F.

HPLC: 10.083 min.

MS: MH⁺=515.1

Example 145-18 Preparation of4-[4-(2,4-dichlorophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-5-yl]morpholine-3,5-dione

4-[4-(2,4-dichlorophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-5-yl]morpholine-3,5-dionewas made in accordance with the foregoing procedures through steps Ausing 1-(2,4-dichlorophenyl)-2-chloroethan-1-one and phthalimide, B, Cusing amino{2-[(5-nitro(2-pyridyl))amino]ethyl}carboxamidine, D, E, Kusing 1,4-dioxane-2,6-dione, and F.

HPLC:

MS: MH⁺=518.1

Example 145-19 Preparation ofN-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]-4-morpholin-4-ylbutanamide

N-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]-4-morpholin-4-ylbutanamidewas made in accordance with the foregoing procedures through steps Ausing 1-(2,4-dichlorophenyl)-2-chloroethan-1-one and phthalimide, B, Cusing amino{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}carboxamidine, D,E, F using 4-chlorobutanoic acid. lmmol ofN-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]-4-bromobutanamide,2 mmol of morpholine, 3 mmol Diisopropylethyl amine, and 0.1 mmol oftetrabutylammonium iodide were stirred at room temperature for fourteenhours. The reaction mixture was concentrated in vacuo and diluted withwater and ethyl acetate. The solution was extracted three times withethyl acetate, dried over sodium sulfate, and purified by columnchromatography with 5-10% methanol in methylenechloride.

HPLC: 4.837 min.

MS: MH⁺=590.2

Example 145-20 Preparation of1-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]piperazine-2,6-dione

1-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]piperazine-2,6-dionewas made in accordance with the foregoing procedures through steps Ausing 1-(2,4-dichlorophenyl)-2-chloroethan-1-one and phthalimide, B, Cusing amino{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}carboxamidine, D,E, F using 2-[(tert-butoxy)-N-(carboxymethyl)carbonylamino]acetic acid,and G.

HPLC: 5.825 min.

MS: MH⁺=532.2

Example 145-21 Preparation of tert-butyl4-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]-3,5-dioxopiperazinecarboxylate

tert-butyl4-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]-3,5-dioxopiperazinecarboxylatewas made in accordance with the foregoing procedures through steps Ausing 1-(2,4-dichlorophenyl)-2-chloroethan-1-one and phthalimide, B, Cusing amino{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}carboxamidine, D,E, and F using 2-[(tert-butoxy)-N-(carboxy-methyl)carbonylamino]aceticacid.

HPLC: 9.137 min.

MS: MH⁺=632.2

Example 145-22 Preparation of tert-butyl4-[4-(2,4-dichlorophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-5-yl]-3,5-dioxopiperazinecarboxylate

tert-butyl4-[4-(2,4-dichlorophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-5-yl]-3,5-dioxopiperazinecarboxylatewas made in accordance with the foregoing procedures through steps Ausing 1-(2,4-dichlorophenyl)-2-chloroethan-1-one and phthalimide, B, Cusing amino{2-[(5-nitro(2-pyridyl))amino]ethyl}carboxamidine, D, E, andF using 2-[(tert-butoxy)-N-(carboxymethyl)carbonylamino]acetic acid.

HPLC: 9.861 min.

MS: MH⁺=617.2

Example 145-23 Preparation of1-[4-(2,4-dichlorophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-5-yl]piperazine-2,6dione

1-[4-(2,4-dichlorophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-5-yl]piperazine-2,6-dionewas made in accordance with the foregoing procedures through steps Ausing 1-(2,4-dichlorophenyl)-2-chloroethan-1-one and phthalimide, B, Cusing amino{2-[(5-nitro(2-pyridyl))amino]ethyl}carboxamidine, D, E, Fusing 2-[(tert-butoxy)-N-(carboxymethyl)carbonylamino]acetic acid, andG.

HPLC: 6.554 min.

MS: MH⁺=517.2

Example 145-24 Preparation of tert-butyl4-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]-2,6-dimethylpiperazinecarboxylate

tert-butyl4-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]-2,6-dimethylpiperazinecarboxylatewas made in accordance with the foregoing procedures through steps Ausing 1-(2,4-dichlorophenyl)-2-chloroethan-1-one and2,6-dimethylpiperazine. 1 mmol of this product, 1 mmol of tert-butyl(tert-butoxycarbonyloxy)formate, and 2 mmol of triethylamine werestirred at room temperature in methylenechloride for four hours. Thereaction mixture was concentrated in vacuo and diluted with water andethyl acetate. The solution was extracted three times with ethylacetate, dried over sodium sulfate, and purified by columnchromatography with 5-10% methanol in methylenechloride. This productwas taken on to steps B and C usingamino{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}carboxamidine.

HPLC: 10.96 min.

MS: MH⁺=632.3

Example 145-25 Preparation of tert-butyl4-[4-(2,4-dichlorophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-5-yl]-2,6-dimethylpiperazinecarboxylate

tert-butyl4-[4-(2,4-dichlorophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-5-yl]-2,6-dimethylpiperazinecarboxylatewas made in accordance with the foregoing procedures through steps Ausing 1-(2,4-dichlorophenyl)-2-chloroethan-1-one and2,6-dimethylpiperazine. 1 mmol of this product, 1 mmol of tert-butyl(tert-butoxycarbonyloxy)formate, and 2 mmol of triethylamine werestirred at room temperature in methylenechloride for four hours. Thereaction mixture was concentrated in vacuo and diluted with water andethyl acetate. The solution was extracted three times with ethylacetate, dried over sodium sulfate, and purified by columnchromatography with 5-10% methanol in methylenechloride. This productwas taken on to steps B and C usingamino{2-[(5-nitro(2-pyridyl))amino]ethyl}carboxamidine.

HPLC: 11.713 min.

MS: MH⁺=617.2

Example 145-26 Preparation of{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-(3,5-dimethylpiperazinyl)pyrimidin-2-yl]amine

{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-(3,5-dimethylpiperazinyl)pyrimidin-2-yl]aminewas made in accordance with the foregoing procedures through steps Ausing 1-(2,4-dichlorophenyl)-2-chloroethan-1-one and2,6-dimethylpiperazine. 1 mmol of this product, 1 mmol of tert-butyl(tert-butoxy-carbonyloxy)formate, and 2 mmol of triethylamine werestirred at room temperature in methylenechloride for four hours. Thereaction mixture was concentrated in vacuo and diluted with water andethyl acetate. The solution was extracted three times with ethylacetate, dried over sodium sulfate, and purified by columnchromatography with 5-10% methanol in methylenechloride. This productwas taken on to steps B, C usingamino{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}carboxamidine, and G.

HPLC: 5.653 min.

MS: MH⁺=532.6

Example 145-27 Preparation of[4-(2,4-dichlorophenyl)-5-(3,5-dimethylpiperazinyl)pyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine

[4-(2,4-dichlorophenyl)-5-(3,5-dimethylpiperazinyl)pyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}aminewas made in accordance with the foregoing procedures through steps Ausing 1-(2,4-dichlorophenyl)-2-chloroethan-1-one and2,6-dimethylpiperazine. 1 mmol of this product, 1 mmol of tert-butyl(tert-butoxycarbonyloxy)formate, and 2 mmol of triethylamine werestirred at room temperature in methylenechloride for four hours. Thereaction mixture was concentrated in vacuo and diluted with water andethyl acetate. The solution was extracted three times with ethylacetate, dried over sodium sulfate, and purified by columnchromatography with 5-10% methanol in methylenechloride. This productwas taken on to steps B, C usingamino{2-[(5-nitro(2-pyridyl))amino]ethyl}carboxamidine, and G.

HPLC: 6.193 min.

MS: MH⁺=517.6

Example 145-28 Preparation ofN-[4-(2,4-dichlorophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-5-yl]-4-hydroxybutanamide

N-[4-(2,4-dichlorophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-5-yl]-4-hydroxybutanamide(74814) was made in accordance with the foregoing procedures throughsteps A using 1-(2,4-dichlorophenyl)-2-chloroethan-1-one andphthalimide, B, C usingamino{2-[(5-nitro(2-pyridyl))amino]ethyl}carboxamidine, D, E, F using4-bromobutanoic acid.

HPLC: 5.688 min.

MS: MH⁺=506.2

Example 145-29 Preparation of[5-((1E)-1-aza-2-pyrrolidinylprop-1-enyl)-4-(2,4-dichlorophenyl)pyrimidin-2-yl]{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amine

[5-((1E)-1-aza-2-pyrrolidinylprop-1-enyl)-4-(2,4-dichlorophenyl)pyrimidin-2-yl]-{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}aminewas made in accordance with the foregoing procedures through steps Ausing 1-(2,4-dichlorophenyl)-2-chloroethan-1-one and phthalimide, B, Cusing amino{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}carboxamnidine,D, E, and K using acetic anhydride. 1 mmol ofN-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]acetamideand 2 mmol of Lawesson's reagent were stirred in 2 ml of DME at 80° C.The reaction mixture was concentrated in vacuo and diluted with waterand ethyl acetate. The solution was extracted three times with ethylacetate, dried over sodium sulfate, and purified by columnchromatography eluting with 5-10% methanol/methylene chloride. 1 mmol of1-{[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]amino}ethane-1-thionewas heated to 85° C. in pyrrolidine and purified by columnchromatography eluting with 5-10% methanol/methylene chloride.

HPLC: 6.032 min.

MS: MH⁺=530.3

Example 145-30 Preparation of{5-[(1E)-1-aza-2-(cyclopropylamino)prop-1-enyl]-4-(2,4-dichlorophenyl)pyrimidin-2-yl}{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amine

{5-[(1E)-1-aza-2-(cyclopropylamino)prop-1-enyl]-4-(2,4-dichlorophenyl)pyrimidin-2-yl}{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}aminewas made in accordance with the foregoing procedures through steps Ausing 1-(2,4-dichlorophenyl)-2-chloroethan-1-one and phthalimide, B, Cusing amino{2-[(6-amnino-5-nitro(2-pyridyl))amino]ethyl}carboxamidine,D, E, and K using acetic anhydride. 1 mmol ofN-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]acetamideand 2 mmol of Lawesson's reagent were stirred in 2 ml of DME at 80° C.The reaction mixture was concentrated in vacuo and diluted with waterand ethyl acetate. The solution was extracted three times with ethylacetate, dried over sodium sulfate, and purified by columnchromatography eluting with 5-10% methanol/methylene chloride. 1 mmol of1-{[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]amino}ethane-1-thionewas heated to 40° C. in cyclopropylamine and purified by columnchromatography eluting with 5-10% methanol/methylene chloride.

HPLC: 5.781 min.

MS: MH⁺=516.2

Example 145-31 Preparation of1-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]-3-(4-methylpiperazinyl)pyrrolidine-2,5-dione

1-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]-3-(4-methylpiperazinyl)pyrrolidine-2,5-dionewas made in accordance with the foregoing procedures through steps Ausing 1-(2,4-dichlorophenyl)-2-chloroethan-1-one and phthalimide, B, Cusing amino{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}carboxamidine, D,E, K using malic anhydride, and F. Large excess of morpholine was addedto clean fractions of1-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]-3-pyrroline-2,5-dioneconcentrated in vacuo, and purified by column chromatography elutingwith 5-10% methanol/methylene chloride.

HPLC: 4.897 min.

MS: MH⁺=546.3

Example 145-32 Preparation of[6-(2,4-dichlorophenyl)-5-nitro(2-pyridyl)]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine

[6-(2,4-dichlorophenyl)-5-nitro(2-pyridyl)]{2-[(5-nitro(2-pyridyl))amino]ethyl}aminewas made in accordance with the foregoing procedures through steps Lusing 2,6-dichloro-3-nitropyridine and 2,4-dichlorobenzeneboronic acid,and step M using (2-aminoethyl)(5-nitro(2-pyridyl))amine.

HPLC: 9.598 min.

MS: MH⁺=448.8

Example 145-33 Preparation ofN-[6-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-2-(2,4-dichlorophenyl)(3-pyridyl)]-N-ethylacetamide

N-[6-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-2-(2,4-dichloro-phenyl)(3-pyridyl)]-N-ethylacetamidewas made in accordance with the foregoing procedures through steps Lusing 2,6-dichloro-3-nitropyridine and 2,4-dichlorobenzeneboronic acid,M using N-(2-aminoethyl)(tert-butoxy)carboxamide, O, K using aceticanhydride, G, and H using 6-chloro-3-nitro-2-pyridylamine.

HPLC: 6.223 min.

MS: MH⁺=504.2

Example 145-34 Preparation of{5-[(6-amino-5-nitro(2-pyridyl))amino]-6-(2,4-dichlorophenyl)(2-pyridyl)}{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amine

{5-[(6-amino-5-nitro(2-pyridyl))amino]-6-(2,4-dichlorophenyl)(2-pyridyl)}{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}aminewas made in accordance with the foregoing procedures through steps Lusing 2,6-dichloro-3-nitropyridine and 2,4-dichlorobenzeneboronic acid,M N-(2-aminoethyl)(tert-butoxy)carboxamide, N, H using6-chloro-3-nitro-2-pyridylamine, G, and H using6-chloro-3-nitro-2-pyridylamine.

HPLC: 7.467 min.

MS: MH⁺=571.0

Example 145-35 Preparation of{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[6-(2,4-dichlorophenyl)-5-(ethylamino)(2-pyridyl)]amine

{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[6-(2,4-dichlorophenyl)-5-(ethylamino)(2-pyridyl)]aminewas made in accordance with the foregoing procedures through steps Lusing 2,6-dichloro-3-nitropyridine and 2,4-dichlorobenzeneboronic acid,M using N-(2-aminoethyl)(tert-butoxy)carboxamide, O, G, and H using6-chloro-3-nitro-2-pyridylamine.

HPLC: 5.263 min.

MS: MH⁺=462.0

Example 145-36 Preparation of[6-(2,4-dichlorophenyl)-3-nitro(2-pyridyl)]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine

[6-(2,4-dichlorophenyl)-3-nitro(2-pyridyl)]{2-[(5-nitro(2-pyridyl))amino]ethyl}aminewas made in accordance with the foregoing procedures through steps Lusing 2,6-dichloro-3-nitropyridine and 2,4-dichlorobenzeneboronic acidcollecting minor product, and step M using(2-aminoethyl)(5-nitro(2-pyridyl))amine.

HPLC: 12.003 min.

MS: MH⁺=449.0

Example 145-37 Preparation of2-(2,4-dichlorophenyl)-4-methyl-6-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyridine-3-carbonitrile

2-(2,4-dichlorophenyl)-4-methyl-6-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyridine-3-carbonitrilewas made in accordance with the foregoing procedures through steps Lusing 2,6-dichloro-4-methylpyridine-3-carbonitrile and2,4-dichloro-benzeneboronic acid, and step M using(2-aminoethyl)(5-nitro(2-pyridyl))amine.

HPLC: 12.183 min.

MS: MH⁺=443.0

Example 145-38 Preparation of{2-[(6-amnino-5-nitro(2-pyridyl))amino]ethyl}[6-(2,4-dichlorophenyl)-3-nitro(2-pyridyl)]amine

{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[6-(2,4-dichlorophenyl)-3-nitro(2-pyridyl)]aminewas made in accordance with the foregoing procedures through steps Lusing 2,6-dichloro-3-nitropyridine and 2,4-dichlorobenzeneboronic acidcollecting minor product, M usingN-(2-aminoethyl)(tert-butoxy)carboxamide, G, and H using6-chloro-3-nitro-2-pyridylamine.

HPLC: 10.682 min.

MS: MH⁺=464.0

Example 145-39 Preparation of{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[6-(4-ethylphenyl)-5-nitro(2-pyridyl)]amine

{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[6-(4-ethylphenyl)-5-nitro(2-pyridyl)]aminewas made in accordance with the foregoing procedures through steps Lusing 2,6-dichloro-3-nitropyridine and 4-ethylbenzeneboronic acid , Musing N-(2-aminoethyl)(tert-butoxy)carboxamide, G, and H using6-chloro-3-nitro-2-pyridylamine.

HPLC: 9.354 min.

MS: MH⁺=424.1

Example 145-40 Preparation ofN-{1-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]-2,6-dioxo(3-piperidyl)}(tert-butoxy)carboxamide

N-{1-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]-2,6-dioxo(3-piperidyl)}(tert-butoxy)carboxamidewas made in accordance with the foregoing procedures through steps Ausing 1-(2,4-dichlorophenyl)-2-chloroethan-1-one and phthalimide, B, Cusing amino{2-[(5-nitro(2-pyridyl))amino]ethyl}carboxamidine, D, E, andF using 1-[(tert-butoxy) carbonyl-amino]propane-1,3-dicarboxylic acid.

HPLC: 9.152 min.

MS: MH⁺=646.4

Example 145-41 Preparation of3-amino-1-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]piperidine-2,6-dione

3-amino-1-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]piperidine-2,6-dionewas made in accordance with the foregoing procedures through steps Ausing 1-(2,4-dichlorophenyl)-2-chloroethan-1-one and phthalimide, B, Cusing amino{2-[(5-nitro(2-pyridyl))amino]ethyl}carboxamidine, D, E, Fusing 1-[(tert-butoxy)carbonylamino]propane-1,3-dicarboxylic acid, andG.

HPLC: 5.247 min.

MS: MH⁺=546.3

Example 145-42 Preparation ofN-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]-2-[(tert-butoxy)-N-methylcarbonylamino]

N-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichloro-phenyl)pyrimidin-5-yl]-2-[(tert-butoxy)-N-methylcarbonylamino]acetamidewas made in accordance with the foregoing procedures through steps Ausing 1-(2,4-dichlorophenyl)-2-chloroethan-1-one and phthalimide, B, Cusing amino{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}carboxamidine, D,E, and F using 2-[(tert-butoxy)-N-methylcarbonylamino]acetic acid.

HPLC: 8.346 min.

MS: MH⁺=606.2

Example 145-43 Preparation ofN-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]-2-(methylamino)acetamide

N-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]-2-(methylamino)acetamidewas made in accordance with the foregoing procedures through steps Ausing 1-(2,4-dichlorophenyl)-2-chloroethan-1-one and phthalimide, B, Cusing amino{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}carboxamidine, D,E, F using 2-[(tert-butoxy)-N-methylcarbonylamino]acetic acid, and G.

HPLC: 4.716 min.

MS: MH⁺=506.1

Example 145-44 Preparation of1-[6-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-2-(4-ethylphenyl)-3-pyridyl]pyrrolidine-2,5-dione

1-[6-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-2-(4-ethylphenyl)-3-pyridyl]pyrrolidine-2,5-dionewas made in accordance with the foregoing procedures through steps Lusing 2,6-dichloro-3-nitropyridine and 4-ethylbenzeneboronic acid , Musing N-(2-aminoethyl)(tert-butoxy)carboxamide, F usingethane-1,2-dicarboxylic acid, G, and H using6-chloro-3-nitro-2-pyridylamine.

HPLC: 6.072 min.

MS: MH⁺=476.2

Example 145-45 Preparation of2-[4-(2,4-dichlorophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-5-yl]isoindoline-1,3-dione

2-[4-(2,4-dichlorophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-5-yl]isoindoline-1,3-dionewas made in accordance with the foregoing procedures through steps Ausing 1-(2,4-dichlorophenyl)-2-chloroethan-1-one and phthalimide, B, Cusing amino{2-[(5-nitro(2-pyridyl))amino]ethyl}carboxamidine, and D.

HPLC: 12.12 min.

MS: MH⁺=549.8

Example 145-46 Preparation of2-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]-3-pyrrolino[3,4-c]pyridine-1,3-dione

2-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]-3-pyrrolino[3,4-c]pyridine-1,3-dionewas made in accordance with the foregoing procedures through steps Ausing 1-(2,4-dichlorophenyl)-2-chloroethan-1-one and3-pyrrolino[3,4-c]pyridine-1,3-dione, B, C usingamino{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}carboxamidine, and D.

HPLC: 9.85 min.

MS: MH⁺=566.1

Example 145-47 Preparation of1-{[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]amino}ethane-1-thione

1-{[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichloro-phenyl)pyrimidin-5-yl]amino}ethane-1-thionewas made in accordance with the foregoing procedures through steps Ausing 1-(2,4-dichlorophenyl)-2-chloroethan-1-one and phthalimide, B, Cusing amino{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}carboxamidine, D,E, and K using acetic anhydride. 1 mmol ofN-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]acetamideand 2 mmol of Lawesson's reagent were stirred in 2 ml of DME at 80° C.The reaction mixture was concentrated in vacuo and diluted with waterand ethyl acetate. The solution was extracted three times with ethylacetate, dried over sodium sulfate, and purified by columnchromatography eluting with 5-10% methanol/methylene chloride.

HPLC: 11.63 min.

MS: MH⁺=493.1

Example 146 Preparation of4-[5-imidazol-2-yl-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]benzenecarbonitrile

4-[5-imidazol-2-yl-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]benzenecarbonitrilewas prepared from 4-cyanobenzoyl chloride using the general method for[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC: 21.9 min (>95% purity)

MS: M+H=428.1 (C₂₁H₁₇N₉O₂+H=428)

Example 147 Preparation of6-[(2-{[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carbonitrile

6-[(2-{[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carbonitrilewas prepared from 2-chloro-5-(cyano)pyridine using the general methodfor[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC: 18.2 min (>95% purity)

MS: M+H=451.1 (C₂₁H₁₆C₁₂N₈+H=451)

Example 148 Preparation of[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl](2-{[5-(trifluoromethyl)(2-pyridyl)]amino}ethyl)amine

[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl](2-{[5-(trifluoromethyl)(2-pyridyl)]amino}ethyl)amine (71480) was prepared from2-chloro-5-(trifluoro-methyl)pyridine using the general method for[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC: 18.9 min (>95% purity)

MS: M+H=494.1 (C₂₁H₁₆Cl₂F₃N₇+H=494)

Example 149 Preparation of[4-(2,4-dichlorophenyl)-5-(1-methylimidazol-2-yl)pyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine

[4-(2,4-dichlorophenyl)-5-(1-methylimidazol-2-yl)pyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}aminewas prepared from 1,2-dimethylimidazole using the general method for[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC: 21.9 min (>95% purity)

MS: M+H=485.1 (C₂₁H₁₈Cl₂N₈O₂+H=485)

Example 150 Preparation of{5-imidazol-2-yl-4-[4-(trifluoromethyl)phenyl]pyrimidin-2-yl}{2-[(5-nitro(2-pyridyl))amino]ethyl}amine

{5-imidazol-2-yl-4-[4-(trifluoromethyl)phenyl]pyrimidin-2-yl}{2-[(5-nitro(2-pyridyl))amino]ethyl}aminewas prepared from 4-(trifluoromethyl)benzoyl chloride using the generalmethod for[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC: 22.0 min (>95% purity)

MS: M+H=471.2 (C₂₁H₁₇F₃N₈O₂+H=471)

Example 151 Preparation of6-{[2-({5-imidazol-2-yl-4-[4-(trifluoromethyl)phenyl]pyrimidin-2-yl}amino)ethyl]amino}pyridine-3-carbonitrile

6-{[2-({5-imidazol-2-yl-4-[4-(trifluoromethyl)phenyl]pyrimidin-2-yl}amino)ethyl]amino}pyridine-3-carbonitrilewas prepared from 4-(trifluoromethyl)benzoyl chloride and2-chloro-5-(cyano)pyridine using the general method for[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC: 19.3 min (>95% purity)

MS: M+H=451.2 (C₂₂H₁₇F₃N₈+H=451)

Example 152 Preparation of{5-imidazol-2-yl-4-[4-(trifluoromethyl)phenyl]pyrimidin-2-yl}(2-{[5-(trifluoromethyl)(2-pyridyl)]amino}ethyl)amine

{5-imidazol-2-yl-4-[4-(trifluoromethyl)phenyl]pyrimidin-2-yl}(2-{[5-(trifluoro-methyl)(2-pyridyl)]amino}ethyl)aminewas prepared from 4-(trifluoromethyl)benzoyl chloride and2-chloro-5-(trifluoromethyl)pyridine using the general method for[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC: 20.0 min (>95% purity)

MS: M+H=494.2 (C₂₂H₁₇F₆N₇+H=494)

Example 153 Preparation of6-[(2-{[4-dichlorophenyl)-5-(1-methylimidazol-2-yl)pyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carbonitrile

6-[(2-{[4-(2,4-dichlorophenyl)-5-(1-methylimidazol-2-yl)pyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carbonitrilewas prepared from 1,2-dimethylimidazole and 2-chloro-5-(cyano)pyridineusing the general method for[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC: 19.0 min (>95% purity)

MS: M+H=465.1 (C₂₂H₁₈Cl₂N₈+H=465)

Example 154 Preparation of[4-(2,4-dichlorophenyl)-5-(1-methylimidazol-2-yl)pyrimidin-2-yl](2-{[5-(trifluoromethyl)(2-pyridyl)]amino}ethyl)amine

[4-(2,4-dichlorophenyl)-5-(1-methylimidazol-2-yl)pyrimidin-2-yl](2-{[5-(trifluoro-methyl)(2-pyridyl)]amino}ethyl)aminewas prepared from 1,2-dimethylimidazole and2-chloro-5-(trifluoromethyl)pyridine using the general method for[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC: 20.0 min (>95% purity)

MS: M+H=508.1 (C₂₂H₁₈Cl₂F₃N₇+H=508)

Example 155 Preparation of[4-(2-chlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine

[4-(2-chlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}aminewas prepared from 2-chlorobenzoyl chloride using the general method for[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC: 18.5 min (>95% purity)

MS: M+H=437.1 (C₂₀H₁₇ClN₈O₂+H=437)

Example 156 Preparation of6-[(2-{[4-(2-chlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carbonitrile

6-[(2-{[4-(2-chlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carbonitrilewas prepared from 2-chlorobenzoyl chloride and2-chloro-5-(cyano)pyridine using the general method for[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC: 15.3 min (>95% purity)

MS: M+H=417.2 (C₂₁H₁₇ClN₈+H=417)

Example 157 Preparation of[4-(2-chlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl](2-{[5-(trifluoromethyl)(2-pyridyl)]amino}ethyl)amine

[4-(2-chlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl](2-{[5-(trifluoromethyl)(2-pyridyl)]amino}ethyl)aminewas prepared from 2-chlorobenzoyl chloride and2-chloro-5-(trifluoromethyl)pyridine using the general method for[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC: 16.8 min (>95% purity)

MS: M+H=460.2 (C₂₁H₁₇CIF₃N₇+H=460)

Example 158 Preparation of[4-(2-chloro-4-fluorophenyl)-5-imidazol-2-ylprimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine

[4-(2-chloro-4-fluorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}aminewas prepared from 2-chloro-4-fluorobenzoyl chloride using the generalmethod for[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC: 19.4 min (>95% purity)

MS: M+H=455.1 (C₂₀H₁₆ClFN₈O₂+H=455)

Example 159 Preparation of{4-[4-fluoro-2-(trifluoromethyl)phenyl]-5-imidazol-2-ylpyrimidin-2-yl}{2-[(5-nitro(2-pyridyl))amino]ethyl}amine

{4-[fluoro-2-(trifluoromethyl)phenyl]-5-imidazol-2-ylpyrimidin-2yl}{2-[(5-nitro(2-pyridyl))amino]ethyl}aminewas prepared from 4-fluoro-2-(trifluoromethyl)benzoyl chloride using thegeneral method for[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC: 21.0 min (>95% purity)

MS: M+H=489.2 (C₂₁H₁₆F₄N₈O₂+H=489)

Example 160 Preparation of{4-[4-fluoro-2-(trifluoromethyl)phenyl]-5-imidazol-2-ylpyrimidin-2-yl}{2-[(5-nitro(2-pyridyl))amino]ethyl}amine

{4-[4-fluoro-2-(trifluoromethyl)phenyl]-5-imidazol-2-ylpyrimidin-2-yl}{2-[(5-nitro(2-pyridyl))amino]ethyl}aminewas prepared from 4-fluoro-2-(trifluoromethyl)benzoyl chloride using thegeneral method for[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC: 21.2 min (>95% purity)

MS: M+H=431.3 (C₂₁H₁₆F₄N₈O₂+H=431)

Example 161 Preparation of[4-(4-ethylphenyl)-5-imidazol-2-ylpyrimidin-2-yl](2-{[5-(trifluoromethyl)(2-pyridyl)]amino}ethyl)amine

[4-(4-ethylphenyl)-5-imidazol-2-ylpyrimidin-2-yl](2-{[5-(trifluoromethyl)(2-pyridyl)]amino}ethyl)aminewas prepared from 4-ethylbenzoyl chloride and2-chloro-5-(trifluoromethyl)pyridine using the general method for[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC: 19.4 min (>95% purity)

MS: M+H=454.3 (C₂₃H₂₂F₃N₇+H=454)

Example 162 Preparation of{4-[2-fluoro-4-(trifluoromethyl)phenyl]-5-imidazol-2-ylpyrimidin-2-yl}{2-[(5-nitro(2-pyridyl))amino]ethyl}amine

{4-[2-fluoro-4-(trifluoromethyl)phenyl]-5-imidazol-2-ylpyrimidin-2-yl}{2-[(5-nitro(2-pyridyl))amino]ethyl}aminewas prepared from 2-fluoro-4-(trifluoromethyl)benzoyl chloride using thegeneral method for[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC: 22.2 min (>95% purity)

MS: M+H=489.2 (C₂₁H₁₆F₄N₈O₂+H=489)

Example 163 Preparation of6-[(2-{[4-(2-chloro-4-fluorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carbonitrile

6-[(2-{[4-(2-chloro-4-fluorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carbonitrilewas prepared from 2-chloro-4-fluorobenzoyl chloride and2-chloro-5-(cyano)pyridine using the general method for[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC: 16.3 min (>95% purity)

MS: M+H=435.2 (C₂₁H₁₆ClFN₈+H=435)

Example 164 Preparation of[4-(2-chloro-4-fluorophenyl)-5-imidazol-2-ylpyrimidin-2-yl](2-{[5-(trifluoromethyl)(2-pyridyl)]amino}ethyl)amine

[4-(2-chloro-4-fluorophenyl)-5-imidazol-2-ylpyrimidin-2-yl](2-{[5-(trifluoromethyl)(2-pyridyl)]amino}ethyl)aminewas prepared from 2-chloro-4-fluorobenzoyl chloride and2-chloro-5-(trifluoromethyl)pyridine using the general method for[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC: 17.7 min (>95% purity)

MS: M+H=478.2 (C₂₁H₁₆ClF₄N₇+H=478)

Example 165 Preparation of6-{[2-({4-[4-fluoro-2-(trifuoromethyl)phenyl]-5-imidazol-2-ylpyrimidin-2-yl}amino)ethyl]amino}pyridine-3-carbonitrile

6-{[2-({4-[4-fluoro-2-(trifluoromethyl)phenyl]-5-imidazol-2-ylpyrimidin-2-yl}amino)ethyl]amino}pyridine-3-carbonitrilewas prepared from 4-fluoro-2-(trifluoromethyl)benzoyl chloride and2-chloro-5-(cyano)pyridine using the general method for[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC: 18.1 min (>95% purity)

MS: M+H=469.2 (C₂₂H₁₆F₄N₈+H=469)

Example 165 Preparation of{4-[4-fluoro-2-(trifluoromethyl)phenyl]-5-imidazol-2-ylpyrimidin-2-yl}(2-{[5-(trifluoromethyl)(2-pyridyl)]amino}ethyl)amine

{4-[4-fluoro-2-(trifluoromethyl)phenyl]-5-imidazol-2-ylpyrimidin-2-yl}(2-{[5-(trifluoromethyl)(2-pyridyl)]amino}ethyl)amine was prepared from4-fluoro-2-(trifluoromethyl)benzoyl chloride and2-chloro-5-(trifluoromethyl)pyridine using the general method for[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC: 18.8 min (>95% purity)

MS: M+H=512.2 (C₂₂H₁₆F₇N₇+H=512)

Example 166 Preparation of6-[(2-{[4-(4-ethylphenyl)-5-imidazol-2-ylpyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carbonitrile

6-[(2-{[4-(4-ethylphenyl)-5-imidazol-2-ylpyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carbonitrilewas prepared from 4-ethylbenzoyl chloride and 2-chloro-5-(cyano)pyridineusing the general method for[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl})amino]ethyl}amine.

HPLC: 17.9 min (>95% purity)

MS: M+H=411.2 (C₂₃H₂₂N₈+H=411)

Example 167 Preparation of[4-(4-chlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine

[4-(4-chlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}aminewas prepared from 4-chlorobenzoyl chloride using the general method for[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC: 20.0 min (>95% purity)

MS: M+H=437.1 (C₂₀H₁₇ClN₈O₂+H=437)

Example 168 Preparation of6-[(2-{[4-(4-chlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carbonitrile

6-[(2-{[4-(4-chlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carbonitrilewas prepared from 4-chlorobenzoyl chloride and2-chloro-5-(cyano)pyridine using the general method for[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC: 17.1 min (>95% purity)

MS: M+H=417.2 (C₂₁H₁₇ClN₈+H=417)

Example 169 Preparation of[4-(4-chloro-2-methylphenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine

[4-(4-chloro-2-methylphenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}aminewas prepared from 4-chloro-2-methylbenzoyl chloride using the generalmethod for[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC: 20.8 min (>95% purity)

MS: M+H=451.2 (C₂₁H₁₉ClN₈O₂+H=451)

Example 170 Preparation of[4-(4-chloro-2-methylphenyl)-5-imidazol-2-ylpyrimidin-2-yl](2-{[5-(trifluoromethyl)(2-pyridyl)]amino}ethyl)amine

[4-(4-chloro-2-methylphenyl)-5-imidazol-2-ylpyrimidin-2-yl](2-{[5-(trifluoro-methyl)(2-pyridyl)]amino}ethyl)aminewas prepared from 4-chloro-2-methylbenzoyl chloride and2-chloro-5-(trifluoromethyl)pyridine using the general method for[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC: 19.2 min (>95% purity)

MS: M+H=474.2 (C₂₂H₁₉ClF₃N₇+H=474)

Example 171 Preparation of6-{[2-({4-[2-fluoro-4-(trifluoromethyl)phenyl]-5-imidazol-2-ylpyrimidin-2-yl}amino)ethyl]amino}pyridine-3-carbonitrile

6-{[2-({4-[2-fluoro-4-(trifluoromethyl)phenyl]-5-imidazol-2-ylpyrimidin-2-yl}amino)ethyl]amino}pyridine-3-carbonitrilewas prepared from 2-fluoro-4-(trifluoromethyl)benzoyl chloride and2-chloro-5-(cyano)pyridine using the general method for[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC: 19.7 min (>95% purity)

MS: M+H=469.3 (C₂₂H₁₆F₄N₈+H=469)

Example 172 Preparation of[4-(2-fluorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine

[4-(2-fluorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}aminewas prepared from 2-fluorbenzoyl chloride using the general method for[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC: 17.9 min (>95% purity)

MS: M+H=421.2 (C₂₀H₁₇FN₈O₂+H=421)

Example 173 Preparation of6-[(2-{[4-(2-fluorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carbonitrile

6-[(2-{[4-(2-fluorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carbonitrilewas prepared from 2-fluorbenzoyl chloride and 2-chloro-5-(cyano)pyridineusing the general method for[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC: 14.7 min (>95% purity)

MS: M+H=401.2 (C₂₁H₁₇FN₈+H=401)

Example 174 Preparation of[4-(4-chloro-2-methoxyphenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine

[4-(4-chloro-2-methoxyphenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}aminewas prepared from 4-chloro-2-methoxybenzoyl chloride using the generalmethod for[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC: 19.9 min (>95% purity)

MS: M+H=467.3 (C₂₁H₁₉ClN₈O₃+H=467)

Example 175 Preparation of6-[(2-{[4-(4-chloro-2-methoxyphenyl)-5-imidazol-2-ylpyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carbonitrile

6-[(2-{[4-(4-chloro-2-methoxyphenyl)-5-imidazol-2-ylpyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carbonitrilewas prepared from 4-chloro-2-methoxybenzoyl chloride and2-chloro-5-(cyano)pyridine using the general method for[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC: 16.9 min (>95% purity)

MS: M+H=447.3 (C₂₂H₁₉ClN₈O+H=447)

Example 176 Preparation of6-[(2-{[4-(4-chloro-2-methylphenyl)-5-imidazol-2-ylpyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carbonitrile

6-[(2-{[4-(2-chloro-4-fluorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carbonitrilewas prepared from 4-chloro-2-methylbenzoyl chloride and2-chloro-5-(cyano)pyridine using the general method for[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC: 17.9 min (>95% purity)

MS: M+H=430.8 (C₂₂H₁₉ClN₈+H=430)

Example 177 Preparation of[4-(4-bromo-2-chlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine

[4-(4-bromo-2-chlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}aminewas prepared from 4-bromo-2-chlorolbenzoyl chloride using the generalmethod for[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC: 21.5 min (>95% purity)

MS: M+H=515.2 (C₂₀H₁₆BrClN₈O₂+H=515)

Example 178 Preparation of6-[(2-{[4-(4-bromo-2-chlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carbonitrile

6-[(2-{[4-(4-bromo-2-chlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carbonitrilewas prepared from 4-bromo-2-chlorobenzoyl chloride and2-chloro-5-(cyano)pyridine using the general method for[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC: 17.7 min (>95% purity)

MS: M+H=495 (C₂₁H₁₆BrClN₈+H=495)

Example 179 Preparation of[4-(4-bromo-2-chlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl](2-{[5-(trifluoromethyl)(2-pyridyl)]amino}ethyl)amine

[4-(4-bromo-2-chlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl](2-{[5-(trifluoro-methyl)(2-pyridyl)]amino}ethyl)aminewas prepared from 4-bromo-2-chlorobenzoyl chloride and2-chloro-5-(trifluoromethyl)pyridine using the general method for[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC: 19.7 min (>95% purity)

MS: M+H=538.2 (C₂₁H₁₆BrClF₃N₇+H=538)

Example 180 Preparation of4-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-5-imidazol-2-ylpyrimidin-4-yl]benzenecarbonitrile

4-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-5-imidazol-2-ylpyrimidin-4-yl]benzenecarbonitrilewas prepared from 4-cyanobenzoyl chloride using the general method for{2-[(4-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amine.

HPLC: 20.0 min (>95% purity)

MS: M+H=443.1 (C₂₁H₁₈N₁₀O₂+H=443)

Example 181 Preparation of{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}{5-imidazol-2-yl-4-[4-(trifluoromethyl)phenyl]pyrimidin-2-yl}amine

{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}{5-imidazol-2-yl-4-[4-(trifluoro-methyl)phenyl]pyrimidin-2-yl}aminewas prepared from 4-(trifluoromethyl)benzoyl chloride using the generalmethod for{2-[(4-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amine.

HPLC: 20.2 min (>95% purity)

MS: M+H=486.2 (C₂₁H₁₈F₃N₉O₂+H=486)

Example 182 Preparation of{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-(1-methylimidazol-2-yl)pyrimidin-2-yl]amine

{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-(1-methyl-imidazol-2-yl)pyrimidin-2-yl]aminewas prepared from 1,2-dimethylimidazole using the general method for{2-[(4-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amine.

HPLC: 19.6 min (>95% purity)

MS: M+H=500.2 (C₂₁H₁₉Cl₂N₉O₂+H=500)

Example 183 Preparation of{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2-chlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amine

{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2-chlorophenyl)-5-imidazol-2-yl-pyrimidin-2-yl]aminewas prepared from 2-chlorobenzoyl chloride using the general method for{2-[(4-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amine.

HPLC: 16.4 min (>95% purity)

MS: M+H=452.7 (C₂₀H₁₈ClN₉O₂+H=452)

Example 184 Preparation of{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2-chloro-4-fluorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amine

{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2-chloro-4-fluorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]aminewas prepared from 2-chloro-4-fluorobenzoyl chloride using the generalmethod for{2-[(4-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amine.

HPLC: 17.3 min (>95% purity)

MS: M+H=470.2 (C₂₀H₁₇ClFN₉O₂+H=470)

Example 185 Preparation of{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}{4-[4-fluoro-2-(trifluoromethyl)phenyl]-5-imidazol-2-ylpyrimidin-2-yl}amine

{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}{4-[4-fluoro-2-(trifluoromethyl)phenyl]-5-imidazol-2-ylpyrimidin-2-yl}aminewas, prepared from 4-fluoro-2-(trifluoromethyl)benzoyl chloride usingthe general method for{2-[(4-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amine.

HPLC: 18.4 min (>95% purity)

MS: M+H=504.3 (C₂₁H₁₇F₄N₉O₂+H=504)

Example 186 Preparation of{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(4-chlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amine

{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(4-chlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]aminewas prepared from 4-chlorobenzoyl chloride using the general method for{2-[(4-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amine.

HPLC: 18.0 min (>95% purity)

MS: M+H=452.2 (C₂₀H₁₈ClN₉O₂+H=452)

Example 187 Preparation of{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(4-chloro-2-methylphenyl)-5-imidazol-2-ylpyrimidin-2-yl]amine

{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(4-chloro-2-methylphenyl)-5-imidazol-2-ylpyrimidin-2-yl]aminewas prepared from 4-chloro-2-methylbenzoyl chloride using the generalmethod for{2-[(4-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amine.

HPLC: 18.7 min (>95% purity)

MS: M+H=466.1 (C₂₁H₂₀CIN₉O₂+H=466)

Example 188 Preparation of{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(4-chloro-2-methoxyphenyl)-5-imidazol-2-ylpyrimidin-2-yl]amine

{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(4-chloro-2-methoxyphenyl)-5-imidazol-2-ylpyrimidin-2-yl]aminewas prepared from 4-chloro-2-methoxybenzoyl chloride using the generalmethod for{2-[(4-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amine.

HPLC: 17.8 min (>95% purity)

MS: M+H=482.1 (C₂₁H₂₀ClN₉O₃+H=482)

Example 189 Preparation of{2-[(6-amino-5-nitro(2-pyridyl)amino]ethyl}[4-(4-bromo-2-chlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amine

{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(4-chloro-2-methoxyphenyl)-5-imidazol-2-ylpyrimidin-2-yl]aminewas prepared from 4-bromo-2-chlorolbenzoyl chloride using the generalmethod for{2-[(4-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amine.

HPLC: 19.4 min (>95% purity)

MS: M+H=530 (C₂₀H₁₇BrClN₉O₂+H=530)

Example 190 Preparation of{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(4-bromo-2-chlorophenyl)-5-(4-methylimidazol-2-yl)pyrimidin-2-yl]amine

{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(4-bromo-2-chlorophenyl)-5-(4-methylimidazol-2-yl)pyrimidin-2-yl]aminewas prepared from 4-bromo-2-chlorolbenzoyl chloride and2-amino-6-chloro-3-nitropyridine using the general method for6-[(2-{[4-(2,4-dichlorophenyl)-5-(4-methylimidazol-2-yl)pyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carbonitrile.

HPLC: 19.4 min (>95% purity)

MS: M+H=544.1 (C₂₁H₁₉BrClN₉O₂+H=544)

Example 191 Preparation of6-[(2-{[4-(4-bromo-2-chlorophenyl)-5-(4-methylimidazol-2-yl)pyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carbonitrile

6-[(2-{[4-(4-bromo-2-chlorophenyl)-5-(4-methylimidazol-2-yl)pyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carbonitrilewas prepared from 4-bromo-2-chlorolbenzoyl chloride using the generalmethod for6-[(2-{[4-(2,4-dichlorophenyl)-5-(4-methylimidazol-2-yl)pyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carbonitrile.

HPLC: 18.7 min (>95% purity)

MS: M+H=509.1 (C₂₂H₁₈BrClN₈+H=509)

Example 192 Preparation of{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2-chloro-4-fluorophenyl)-5-(4-methylimidazol-2-yl)pyrimidin-2-yl]amine

{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2-chloro-4-fluorophenyl)-5-(4-methylimidazol-2-yl)pyrimidin-2-yl]aminewas prepared from 4-fluoro-2-chlorobenzoyl chloride and2-amino-6-chloro-3-nitropyridine using the general method for6-[(2-{[4-(2,4-dichlorophenyl)-5-(4-methylimidazol-2-yl)pyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carbonitrile.

HPLC: 17.7 min (>95% purity)

MS: M+H=484.3 (C₂₁H₁₉ClFN₉O₂+H=484)

Example 193 Preparation of6-[(2-{[4-(2-chloro-4-fluorophenyl)-5-(4-methylimidazol-2-yl)pyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carbonitrile

6-[(2-{[4-(2-chloro-4-fluorophenyl)-5-(4-methylimidazol-2-yl)pyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carbonitrilewas prepared from 4-fluoro-2-chlorobenzoyl chloride using the generalmethod for6-[(2-{[4-(2,4-dichlorophenyl)-5-(4-methylimidazol-2-yl)pyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carbonitrile.

HPLC: 16.7 min (>95% purity)

MS: M+H=449.3 (C₂₂H₁₈ClFN₈+H=449)

Example 194 Preparation of[4-(2,4-dichlorophenyl)-5-(4-methylimidazol-2-yl)pyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine

[4-(2,4-dichlorophenyl)-5-(4-methylimidazol-2-yl)pyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}aminewas prepared from 6-chloro-3-nitropyridine using the general method for6-[(2-{[4-(2,4-dichlorophenyl)-5-(4-methylimidazol-2-yl)pyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carbonitrile.

HPLC: 21.9 min (>95% purity)

MS: M+H=485.6 (C₂₁H₁₈Cl₂N₈O₂+H=485)

Example 195 Preparation of{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-(4-methylimidazol-2-yl)pyrimidin-2-yl]amine

{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-(4-methyl-imidazol-2-yl)pyrimidin-2-yl]aminewas prepared from 2-amino-6-chloro-3-nitropyridine using the generalmethod for6-[(2-{[4-(2,4-dichlorophenyl)-5-(4-methylimidazol-2-yl)pyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carbonitrile.

HPLC: 19.8 min (>95% purity)

MS: M+H=500.2 (C₂₁H₁₉C₁₂N₉O₂+H=500)

Example 196 Preparation of[4-(2,4-dichlorophenyl)-5-(4-methylimidazol-2-yl)pyrimidin-2-yl](2-{[5-(trifluoromethyl)(2-pyridyl)]amino}ethyl)amine

[4-(2,4-dichlorophenyl)-5-(4-methylimidazol-2-yl)pyrimidin-2-yl](2-{[5-(trifluoro-methyl)(2-pyridyl)]amino}ethyl)aminewas prepared from 2-chloro-5-(trifluoromethyl)pyridine using the generalmethod for6-[(2-{[4-(2,4-dichlorophenyl)-5-(4-methylimidazol-2-yl)pyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carbonitrile.

HPLC: 20.0 min (>95% purity)

MS: M+H=508.1 (C₂₂H₁₈Cl₂F₃N₇+H=508)

Example 197 Preparation of4-[2-({2-[(6-chloropyrimidin-4-yl)amino]ethyl}amino)-5-imidazolylpyrimidin-4-yl]benzenecarbonitrile

4-[2-({2-[(6-chloropyrimidin-4-yl)amino]ethyl}amino)-5-imidazolylpyrimidin-4-yl]benzenecarbonitrilewas prepared from 4,6-dichloropyrimidine using the general method for4-{5-imidazolyl-2-[(2-{[5-(trifluoromethyl)(2-pyridyl)]amino}ethyl)amino]pyrimidin-4-yl}benzenecarbonitrile.

HPLC: 16.1 min (>95% purity)

MS: M+H=418.1 (C₂₀H₁₆ClN₉+H=418)

Example 198 Preparation of4-amino-2-[(2-{[4-(4-cyanophenyl)-5-imidazolylpyrimidin-2-yl]amino}ethyl)amino]pyrimidine-5-carbonitrile

4-amino-2-[(2-{[4-(4-cyanophenyl)-5-imidazolylpyrimidin-2-yl]amino}ethyl)amino]pyrimidine-5-carbonitrilewas prepared from 4-amino-2-chloropyrimidine-5-carbonitrile using thegeneral method for 4-{5-imidazolyl-2-[(2-{[5-(trifluoromethyl)(2-pyridyl)]amino}ethyl)amino]pyrimidin-4-yl}benzenecarbonitrile.

HPLC: 17.5 min (>95% purity)

MS: M+H=436.2 (C₂₁H₁₇N₁₁+H=436)

Example 199 Preparation of[6-(2,4-dichlorophenyl)-5-(4-methylimidazolyl)(2-pyridyl)]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine

[6-(2,4-dichlorophenyl)-5-(4-methylimidazolyl)(2-pyridyl)]{2-[(5-nitro(2-pyridyl))amino]ethyl}aminewas prepared from 4-methyimidazole (the isomers were separated using aslica gel column) and 2-chloro-5-nitropyridine using the general methodfor{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[6-(2,4-dichlorophenyl)-5-imidazolyl(2-pyridyl)]amine.

HPLC: 22.8 min (>95% purity)

MS: M+H=484.2 (C₂₂H₁₉Cl₂N₇O₂+H=484)

Example 200 Preparation of[6-(2,4-dichlorophenyl)-5-(4-methylimidazolyl)(2-pyridyl)](2-{[5-(trifluoromethyl)(2-pyridyl)]amino}ethyl)amine

[6-(2,4-dichlorophenyl)-5-(4-methylimidazolyl)(2-pyridyl)](2-{[5-(trifluoro-methyl)(2-pyridyl)]amino}ethyl)aminewas prepared from 4-methyimidazole (the isomers were separated using aslica gel column) and 2-chloro-5-(trifluoromethyl)pyridine using thegeneral method for{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[6-(2,4-dichlorophenyl)-5-imidazolyl(2-pyridyl)]amine.

HPLC: 21.0 min (>95% purity)

MS: M+H=507 (C₂₃H₁₉Cl₂F₃N₆+H=507)

Example 201 Preparation of1-[2-(2,4-dichlorophenyl)-6-({2-pyridyl))amino]ethyl}amino)-3-pyridyl]hydropyridin-2-one

1-[2-(2,4-dichlorophenyl)-6-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-3-pyridyl]hydropyridin-2-onewas prepared following the same procedures as the method for{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[6-(2,4-dichlorophenyl)-5-imidazolyl(2-pyridyl)]amine with the following exceptions.1-[2-(2,4-dichlorophenyl)-2-oxoethyl]hydropyridin-2-one was made byheating 2-hydroxypyridine with 2 equivalents of Hünig's base inacetonitrile until dissolved followed by addition of1-(2,4-dichlorophenyl)-2-chloroethan-1-one. The reaction was heated at65° C. for 18 hours and purified by slica gel column. Also,2-chloro-5-nitropyridine was used for the final step.

HPLC: X min (>95% purity)

MS: M+H=X (C₂₃H₁₈Cl₂N₆O₃+H=X)

Example 202 Preparation of1-[6-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-2-(2,4-dichlorophenyl)-3-pyridyl]hydropyridin-2-one

1-[6-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-2-(2,4-dichlorophenyl)-3-pyridyl]hydropyridin-2-onewas prepared following the same procedures as the method for{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[6-(2,4-dichlorophenyl)-5-imidazolyl(2-pyridyl)]aminewith the following exceptions.1-[2-(2,4-dichlorophenyl)-2-oxoethyl]hydropyridin-2-one was made byheating 2-hydroxypyridine with 2 equivalents of Hünig's base inacetonitrile until dissolved followed by addition of1-(2,4-dichlorophenyl)-2-chloroethan-1-one. The reaction was heated at65° C. for 18 hours and purified by slica gel column.

HPLC: X min (>95% purity)

MS: M+H=X (C₂₃H₁₉Cl₂N₇O₃+H=X)

Example 203 Preparation of6-[(2-{[6-(2,4-dichlorophenyl)-5-(2-oxohydropyridyl)-2-pyridyl]amino}ethyl)amino]pyridine-3-carbonitrile

6-[(2-{[6-(2,4-dichlorophenyl)-5-(2-oxohydropyridyl)-2-pyridyl]amino}ethyl)amino]pyridine-3-carbonitrilewas prepared following the same procedures as the method for{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[6-(2,4-dichlorophenyl)-5-imidazolyl(2-pyridyl)]aminewith the following exceptions.1-[2-(2,4-dichlorophenyl)-2-oxoethyl]hydropyridin-2-one was made byheating 2-hydroxypyridine with 2 equivalents of Hünig's base inacetonitrile until dissolved followed by addition of1-(2,4-dichlorophenyl)-2-chloroethan-1-one. The reaction was heated at65° C. for 18 hours and purified by slica gel column. Also,2-chloro-5-cyanopyridine was used for the final step.

HPLC: X min (>95% purity)

MS: M+H=X (C₂₄H₁₈Cl₂N₆O+H=X)

Example 204 Preparation of ethyl6-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-2-phenylpyridine-3-carboxylate

ethyl6-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-2-phenylpyridine-3-carboxylatewas prepared from ethyl 3-oxo-3-phenylpropanoate as the startingmaterial and DDQ in the oxidation step. The final product is achieveddirectly from the chloropyridine by reacting ethyl6-chloro-2-phenylpyridine-3-carboxylate with2-(2-aminoethylamine)-5-nitropyridine in CH₃CN and Hüing's base at 80°C. for 18 hours. The procedures are analogous to the general method for{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[6-(2,4-dichlorophenyl)-5-imidazolyl(2-pyridyl)]amine.

HPLC: 24.5 min (>95% purity)

MS: M+H=408.1 (C₂₁H₂₁N₅O₄+H=408.1)

Example 205 Preparation of ethyl2-(2,4-dichlorophenyl)-6-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyridine-3-carboxylate

Ethyl2-(2,4-dichlorophenyl)-6-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyridine-3-carboxylatewas prepared from ethyl 3-(2,4-dichlorophenyl)-3-oxopropanoate (ref.Wemple, J.; et. al. Synthesis 1993, 290-292.) as the starting materialand THF/ethanol ratio of 3:1 as the solvent in the first step. Theoxidation uses DDQ. The final product is achieved directly from thechloropyridine by reacting ethyl 6-chloro-2-phenylpyridine-3-carboxylatewith 2-(2-aminoethylarnine)-5-nitropyridine in CH₃CN and Hüing's Hüing'sbase at 120° C. for 18 hours. The procedures are analogous to thegeneral method for{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[6-(2,4-dichlorophenyl)-5-imidazolyl(2-pyridyl)]amine.

HPLC: 31 min (>95% purity)

MS: M+H=476.1 (C₂₁H₁₉Cl₂N₅O₄+H=476)

Example 206 Preparation of ethyl2-(4-cyanophenyl)-6-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyridine-3-carboxylate

ethyl2-(4-cyanophenyl)-6-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyridine-3-carboxylate(62258) was prepared from ethyl 3-(4-cyanophenyl)-3-oxopropanoate (ref.Wemple, J.; et. al. Synthesis 1993, 290-292.) as the starting materialand THF/ethanol ratio of 1:5 as the solvent in the first step. Theoxidation uses DDQ in toluene. The final product is achieved directlyfrom the chloropyridine by reacting ethyl6-chloro-2-phenylpyridine-3-carboxylate with2-(2-aminoethylamine)-5-nitropyridine in DMA and Hüing's base at 120° C.for 18 hours. The procedures are analogous to the general method for{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[6-(2,4-dichlorophenyl)-5-imidazolyl(2-pyridyl)]amine.

HPLC: 26.8 min (>95% purity)

MS: M+H=433.1 (C₂₂H₂₀N₆O₄+H=433)

Example 207 Preparation of4-[3-imidazolyl-6-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-2-pyridyl]benzenecarbonitrile

4-[3-imidazolyl-6-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-2-pyridyl]benzene-carbonitrilewas prepared from 4-cyanophenacyl bromide, CAN for the oxidation (1:1acetic acid and water with heating at 80° C. for 1 hour), and2-chloro-5-nitropyridine using the general method for{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[6-(2,4-dichlorophenyl)-5-imidazolyl(2-pyridyl)]amine.

HPLC: 19.5 min (>95% purity)

MS: M+H=427.2 (C₂₂H₁₈N₈O₂+H=427)

Example 208 Preparation of ethyl6-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-2-[4-(trifluoromethyl)phenyl]pyridine-3-carboxylate

ethyl6-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-2-[4-(trifluoromethyl)phenyl]pyridine-3-carboxylate was prepared from ethyl3-oxo-3[4-(trifluoromethyl)phenyl]propanoate (ref. Wemple, J.; et. al.Synthesis 1993, 290-292.) as the starting material. The oxidation wasaccomplished using 4 equivalents of chlorotrimethysilane and 1equivalent of bromine in dichlormethane. The product was obtaineddirectly from ethyl6-chloro-2-[4-(trifluoromethyl)phenyl]pyridine-3-carboxylate by reactingit with 2-(2-aminoethylamine)-5-nitropyridine in DMA and Hünig's base at70° C. for 72 hours. These procedures are analogous to the generalmethod for{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[6-(2,4-dichlorophenyl)-5-imidazolyl(2-pyridyl)]amine.

HPLC: 30.4 min (>95% purity)

MS: M+H=476.2 (C₂₂H₂₀F₃N₅O₄+H=476)

Example 209 Preparation of6-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-2-[4-(trifluoromethyl)phenyl]pyridine-3-carboxylicacid

6-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-2-[4-(trifluoromethyl)phenyl]pyridine-3-carboxylic acid was made by hydrolyzing ethyl6-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-2-[4-(trifluoromethyl)phenyl]pyridine-3-carboxylate(71477) which was made following the general method for{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[6-(2,4-dichlorophenyl)-5-imidazolyl(2-pyridyl)]amine.The hydrolyze was carried out using a solution of 1:1 water andconcentrated hydrochloric acid and heating to 80° C. overnight.

HPLC: 24.0 min (>95% purity)

MS: M+H=448.1 (C₂₀H₁₆F₃N₅O₄+H=448)

Example 210 Preparation of2-(2,4-dichlorophenyl)-6-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyridine-3-carboxylicacid

2-(2,4-dichlorophenyl)-6-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyridine-3-carboxylicacid was made by hydrolyzing ethyl2-(2,4-dichlorophenyl)-6-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyridine-3-carboxylate(62257) which was made following the general method for{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[6-(2,4-dichlorophenyl)-5-imidazolyl(2-pyridyl)]amine.The hydrolize was carried out using a solution of 1:1 water andconcentrated hydrochloric acid and heating to 80° C. overnight.

HPLC: 23.6 min (>95% purity)

MS: M+H=448.1 (C₁₉H₁₅Cl₂N₅O₄+H=448)

Example 211 Preparation of[6-(2,4-dichlorophenyl)-5-imidazolyl(2-pyridyl)]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine

[6-(2,4-dichlorophenyl)-5-imidazolyl(2-pyridyl)]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine was prepared from 2-chloro-5-(nitro)pyridine using thegeneral method for{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[6-(2,4-dichlorophenyl)-5-imidazolyl(2-pyridyl)]amine.

HPLC: 22.9 min (>95% purity)

MS: M+H=470.1 (C₂₁H₁₇Cl₂N₇O₂+H=470)

Example 212 Preparation of6-[(2-{[6-(2,4-dichlorophenyl)-5-imidazolyl-2-pyridyl]amino}ethyl)amino]pyridine-3-carbonitrile

6-[(2-{[6-(2,4-dichlorophenyl)-5-imidazolyl-2-pyridyl]amino}ethyl)amino]pyridine-3-carbonitrilewas prepared from 2-chloropyridine-5-carbonitrile using the generalmethod for{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[6-(2,4-dichlorophenyl)-5-imidazolyl(2-pyridyl)]amine.

HPLC: 18.8 min (>95% purity)

MS: M+H=450 (C₂₂H₁₇Cl₂N₇+H=450)

Example 213 Preparation of{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[6-(2,4-dichlorophenyl)-5-(4-methylimidazolyl)(2-pyridyl)]amine

{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[6-(2,4-dichlorophenyl)-5-(4-methyl-imidazolyl)(2-pyridyl)]aminewas prepared from 4-methyimidazole (the isomers were separated using aslica gel column) using the general method for{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[6-(2,4-dichlorophenyl)-5-imidazolyl(2-pyridyl)]amine.

HPLC: 21.5 min (>95% purity)

MS: M+H=499.3 (C₂₂H₂₀Cl₂N₈O₂+H=499)

Example 214 Preparation of6-[(2-{[6-(2,4-dichlorophenyl)-5-(4-methylimidazolyl)-2-pyridyl]amino}ethyl)amino]pyridine-3-carbonitrile

6-[(2-{[6-(2,4-dichlorophenyl)-5-(4-methylimidazolyl)-2-pyridyl]amino}ethyl)amino]pyridine-3-carbonitrilewas prepared from 4-methyimidazole (The isomers were separated using aslica gel column.) and 2-chloropyridine-5-carbonitrile using the generalmethod for{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[6-(2,4-dichlorophenyl)-5-imidazolyl(2-pyridyl)]amine.

HPLC: 20.7 min (>95% purity)

MS: M+H=464.2 (C₂₃H₁₉Cl₂N₇+H=464)

Example 215 Preparation of{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[6-(2,4-dichlorophenyl)-5-imidazol-2-yl(2-pyridyl)]amine

{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[6-(2,4-dichlorophenyl)-5-imidazol-2-yl(2-pyridyl)]aminewas prepared from (2-aminoethyl)(6-amino-5-nitro(2-pyridyl))amine usingthe general method for[6-(2,4-dichlorophenyl)-5-imidazol-2-yl(2-pyridyl)]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC: 20.1 min (>95% purity)

MS: M+H=485.4 (C₂₁H₁₈Cl₂N₈O₂+H=485)

Example 216 Preparation of[6-(2,4-dichlorophenyl)-5-imidazol-2-yl(2-pyridyl)](2-{[5-(trifluoromethyl)(2-pyridyl)]amino}ethyl)amine

[6-(2,4-dichlorophenyl)-5-imidazol-2-yl(2-pyridyl)](2-{[5-(trifluoromethyl)(2-pyridyl)]amino}ethyl)amine was prepared from(2-aminoethyl)[5-(trifluoromethyl) (2-pyridyl)]amine using the generalmethod for[6-(2,4-dichlorophenyl)-5-imidazol-2-yl(2-pyridyl)]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC: 20.3 min (>95% purity)

MS: M+H=493.3 (C₂₂H₁₇Cl₂F₃N₆+H=493)

Example 217 Preparation of6-[(2-{[6-(2,4-dichlorophenyl)-5-imidazol-2-yl-2-pyridyl]amino}ethyl)amino]pyridine-3-carbonetrile

6-[(2-{[6-(2,4-dichlorophenyl)-5-imidazol-2-yl-2-pyridyl]amino}ethyl)amino]pyridine-3-carbonitrilewas prepared from 6-[(2-aminoethyl)amino]pyridine-3-carbonitrile usingthe general method for[6-(2,4-dichlorophenyl)-5-imidazol-2-yl(2-pyridyl)]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine.

HPLC: 18.9 min (>95% purity)

MS: M+H=450.4 (C₂₂H₁₇Cl₂N₇+H=450)

Example 218 Preparation of 1-(2,4-Dichlorophenyl)-2-pyrazolylethan-1-one

To a solution of 2′,4′-dichlorophenacyl chloride (2.0 g, 8.9 mmol) anddry MeCN (50 mL) at 23° C. was added pyrazole (3.1 g, 44.8 mmol). Theresulting solution was heated at 80° C. for 5 h then cooled to 23° C.The MeCN was removed under reduced pressure and CH2Cl2 (50 mL) wasadded. The resulting solution was washed with H2O (2×15 mL) and theorganic layer was dried (Na2SO4) and concentrated under reducedpressure. The resulting residue was purified on silica gel (40%EtOAc/hexanes) to yield a light yellow solid.

m/z 256 (MH+)

Example 219 Preparation of1-(2,4-Dichlorophenyl)-2-(4-methylimidazolyl)ethan-1-one

Made using the same procedure as for1-(2,4-dichlorophenyl)-2-pyrazolylethan-1-one except that4-methylimidazole,(3.7 g, 44.8 mmol) was used in place of pyrazole. Thecrude residue was purified on silica gel (5% MeOH/CH2Cl2) to yield alight yellow solid.

m/z 270 (MH+).

Example 220 Preparation of1-(2,4-Dichlorophenyl)-2-(2,4-dimethylimidazole)ethan-1-one

Made using the same procedure as for1-(2,4-dichlorophenyl)-2-pyrazolylethan-1-one except that2,4-dimethylimidazole (4.3 g, 44.8 mmol) was used in place of pyrazole.The crude residue was purified on silica gel (5% MeOH/CH2Cl2) to yield alight yellow solid

m/z 284 (MH+)

Example 221 Preparation of1-[2-(2,4-Dichlorophenyl)-2-oxoethyl]hydropyridin-2-one

To a solution of 2′, 4′-dichlorophenacyl chloride (1.0 g, 4.5 mmol) anddry MeCN (20 mL) at 23° C. was added polystyrene-bound1,5,7-triazabicyclo[4.4.0]dec-5-ene (2.6 g, 6.7 mmol) and2-hydroxypyridine (428 mg, 4.5 mmol) and the resulting mixture wasshaken at 23° C. for 20 h. The mixture was filtered and the resin waswashed with MeCN (10 mL). The MeCN was removed under reduced pressureand the resulting residue was purified on silica gel (5% MeOH/CH2Cl2) toyield a light yellow solid.

m/z 283 (MH+)

Example 222 Preparation of2-Benzimidazolyl-1-(2,4-dichlorophenyl)ethan-1-one

Made using the same procedure as for1-(2,4-dichlorophenyl)-2-pyrazolylethan-1-one except that benzimidazole(5.3 g, 44.8 mmol) was used in place of pyrazole. The crude residue waspurified on silica gel (50% EtOAc/hexanes) to yield a light yellow solid

m/z 306 (MH+)

Example 223 Preparation of1-(2,4-Dichlorophenyl)-2-(2-methylimidazolyl)ethan-1-one

Made using the same procedure as for1-(2,4-dichlorophenyl)-2-pyrazolylethan-1-one except that2-methylimidazole (3.7 g, 44.8 mmol) was used in place of pyrazole. Thecrude residue was purified on silica gel (5% MeOH/CH2Cl2) to yield alight yellow solid.

m/z 270 (MH+)

Example 224 Preparation of1-(2,4-Dichlorophenyl)-2-(4-phenylimidazolyl)ethan-1-one

Made using the same procedure as for1-(2,4-dichlorophenyl)-2-pyrazolylethan-1-one except that4-phenylimidazole (6.5 g, 44.8 mmol) was used in place of pyrazole. Thecrude residue was purified on silica gel (50% EtOAc/hexanes) to yield alight yellow solid. m/z 332 (MH+)

Example 225 Preparation of1-(2,4-Dichlorophenyl)-2-imidazolylethan-1-one

Made using the same procedure as for1-(2,4-dichlorophenyl)-2-pyrazolylethan-1-one except that imidazole (3.1g, 44.8 mmol) was used in place of pyrazole. The crude residue waspurified on silica gel (5% MeOH/CH2Cl2) to yield a light yellow solid.

m/z 256 (MH+)

Example 226 Preparation of1-[2-(2,4-Dichlorophenyl)-2-oxoethyl]-5-chlorohydropyridin-2-one

Made using the same procedure as for1-[2-(2,4-dichlorophenyl)-2-oxoethyl]hydropyridin-2-one except that5-chloro-2-hydroxypyridine (583 mg, 4.5 mmol) was used in place of2-hydroxypyridine. The crude residue was purified on silica gel (5%MeOH/CH2Cl2) to yield a light yellow solid.

m/z 317 (MH+)

Example 227 Preparation of1-(2,4-Dichlorophenyl)-3-(dimethylamino)-2-pyrazolylprop-2-en-1-one

A solution of 1-(2,4-dichlorophenyl)-2-pyrazolylethan-1-one (I, 1.0 g,3.9 mmol) and dimethylfornamidedimethyl acetal (10 mL, 75 mmol) washeated at 100° C. for 2 h. The resulting red-brown solution wasconcentrated under reduced pressure to yield a dark red-brown oil whichwas used without further purification.

m/z 311 (MH+)

Example 228 Preparation of1-(2,4-Dichlorophenyl)-3-(dimethylamino)-2-(4-methylimidazolyl)prop-2-en-1-one

Made using the same procedure as for1-(2,4-dichlorophenyl)-3-(dimethylamino)-2-pyrazolylprop-2-en-1-oneexcept that 1-(2,4-dichlorophenyl)-2-(4-methylimidazolyl) ethan-1-one(II, 1.0 g, 3.7 mmol) was used. The crude residue was used withoutpurification.

m/z 325 (MH+)

Example 229 Preparation of1-(2,4-Dichlorophenyl)-3-(dimethylamino)-2-(2,4-dimethylimidazolyl)prop-2-en-1-one

Made using the same procedure as for1-(2,4-dichlorophenyl)-3-(dimethylamino)-2-pyrazolylprop-2-en-1-oneexcept that 1-(2,4-dichlorophenyl)-2-(2,4-dimethylimidazole)ethan-1-one(III, 1.0 g, 3.5 mmol) was used. The crude residue was used withoutpurification.

m/z 339 (MH+)

Example 230 Preparation of1-(2,4-Dichlorophenyl)-3-(dimethylamino)-2-(2-oxohydropyridyl)prop-2-en-1-one

A solution of 1-[2-(2,4-dichlorophenyl)-2-oxoethyl]hydropyridin-2-one(IV, 1.0 g, 3.5 mmol), dry THF (25 mL) and dimethylformamidedimethylacetal (10 mL, 75 mmol) was heated at 75° C. for 2.5 h. The resultingdark solution was concentrated under reduced pressure and the oilyresidue was used without purification.

m/z 338 (MH+)

Example 231 Preparation of2-Benzimidazolyl-1-(2,4-dichlorophenyl)-3-(dimethylamino)prop-2-en-1-one

Made using the same procedure as for1-(2,4-dichlorophenyl)-3-(dimethylamino)-2-pyrazolylprop-2-en-1-oneexcept that 2-benzimidazolyl-1-(2,4-dichlorophenyl)ethan-1-one (V, 1.0g, 3.3 mmol) was used. The crude residue was used without purification.

m/z 361 (MH+)

Example 232 Preparation of1-(2,4-Dichlorophenyl)-3-(dimethylamino)-2-(2-methylimidazolyl)prop-2-en-1-one

Made using the same procedure as for1-(2,4-dichlorophenyl)-3-(dimethylamino)-2-pyrazolylprop-2-en-1-oneexcept that 1-(2,4-dichlorophenyl)-2-(2-methylimidazolyl) ethan-1-one(VI, 1.0 g, 3.7 mmol) was used. The crude residue was used withoutpurification.

m/z 325 (MH+)

Example 233 Preparation of1-(2,4-Dichlorophenyl)-3-(dimethylamino)-2-(4-phenylimidazolyl)prop-2-en-1-one

Made using the same procedure as for1-(2,4-dichlorophenyl)-3-(dimethylamino)-2-(2-oxohydropyridyl)prop-2-en-1-oneexcept that 1-(2,4-dichlorophenyl)-2-(4-phenyl-imidazolyl)ethan-1-one(VII, 1.0 g, 3.0 mmol) was used. The crude residue was used withoutpurification.

m/z 387 (MH+)

MS:

Example 234 Preparation of1-(2,4-Dichlorophenyl)-3-(dimethylamino)-2-imidazolylprop-2-en-1-one

Made using the same procedure as for1-(2,4-dichlorophenyl)-3-(dimethylamino)-2-(2-oxohydropyridyl)prop-2-en-1-oneexcept that 1-(2,4-dichlorophenyl)-2-imidazolylethan-1-one (VIII, 1.0 g,3.9 mmol) was used. The crude residue was used without purification.

m/z 311 (MH+)

Example 235 Preparation of1-{2-(2,4-Dichlorophenyl)-1-[(dimethylamino)methylene]-2-oxoethyl}-5-chlorohydropyridin-2-one

Made using the samne procedure as for1-(2,4-dichlorophenyl)-3-(dimethylamino)-2-(2-oxohydropyridyl)prop-2-en-1-oneexcept that1-[2-(2,4-dichlorophenyl)-2-oxoethyl]-5-chlorohydropyridin-2-one (IX,1.0 g, 3.2 mmol) was used. The crude residue was used withoutpurification.

m/z 372 (MH+)

Example 236 Preparation of[4-(2,4-Dichlorophenyl)-5-pyrazolylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}aminedihydrochloride

To a solution of1-(2,4-dichlorophenyl)-3-(dimethylamino)-2-pyrazolylprop-2-en-1-one (X,400 mg, 1.3 mmol) and EtOH (10 mL) at 23° C. was addedamino{2-[(5-nitro(2-pyridyl))amino]ethyl}carboxamidine hydrochloride(365 mg, 1.4 mmol) followed by NaOEt in EtOH (1.6 mL, 1.6 mmol) and theresulting solution was heated at 90° C. for 16 h. The EtOH was removedunder reduced pressure and the resulting residue was purified on silicagel (1-5% MeOH/CH2Cl2) to yield a yellow solid which was dissolved inMeCN/0.5M HCl (3 mL, 1:1), frozen and lyophilized to yield a yellowsolid.

m/z 472 (MH+)

Example 237 Preparation of[4-(2,4-Dichlorophenyl)-5-(4-methylimidazolyl)pyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}aminedihydrochloride

Made using the same procedure as for[4-(2,4-dichlorophenyl)-5-pyrazolylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}aminedihydrochloride except that1-(2,4-dichlorophenyl)-3-(dimethylamino)-2-(4-methylimidazolyl)prop-2-en-1-one(XI, 421 mg, 1.3 mmol) was used.

m/z 486 (MH+)

Example 238 Preparation of[4-(2,4-Dichlorophenyl)-5-(2,4-dimethylimidazolyl)pyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}aminedihydrochloride

Made using the same procedure as for XIX except that1-(2,4-dichlorophenyl)-3-(dimethylamino)-2-(2,4-dimethylimidazolyl)prop-2-en-1-one(XII, 439 mg, 1.3 mmol) was used.

m/z 500 (MH+)

Example 239 Preparation of1-[4-(2,4-Dichlorophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-5-yl]hydropyridin-2-onehydrochloride

Made using the same procedure as for[4-(2,4-dichlorophenyl)-5-pyrazolylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}aminedihydrochloride except that1-(2,4-dichlorophenyl)-3-(dimethylamino)-2-(2-oxohydropyridyl)prop-2-en-1-one(438 mg, 1.3 mmol) was used and the crude product was purified byrecrystallization (CH2Cl2/Et2O/hexanes)

m/z499 (MH+)

Example 240 Preparation of[5-Benzimidazolyl-4-(2,4-dichlorophenyl)pyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}aminedihydrochloride

Made using the same procedure as for[4-(2,4-dichlorophenyl)-5-pyrazolylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amninedihydro chloride except that2-benzimidazolyl-1-(2,4-dicelorophenyl)-3-(dimethylamino)prop-2-en-1-one(XIV, 468 mg, 1.3 mmol) was used and the crude product was purified byrecrystallization (CH₂Cl₂/Et₂O/hexanes).

m/z 522 (MH+)

Example 241 Preparation of[4-(2,4-Dichlorophenyl)-5-(2-methylimidazolyl)pyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}aminedihydrochloride

Made using the same procedure as for[4-(2,4-dichlorophenyl)-5-pyrazolylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}aminedihydrochloride except that1-(2,4-dichlorophenyl)-3-(dimethylamino)-2-(2-methylimidazolyl)prop-2-en-1-one(XV, 421 mg, 1.3 mmol) was used. m/z 486 (MH+)

Example 242 Preparation of{2-[(6-Amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-(2-methylimidazolyl)pyrimidin-2-yl]aminedihydrochloride

Made using the same procedure as for[4-(2,4-dichlorophenyl)-5-pyrazolylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}aminedihydrochloride except that1-(2,4-dichlorophenyl)-3-(dimethylamino)-2-(2-methylimidazolyl)prop-2-en-1-one(XV, 421 mg, 1.3 mmol) andamino{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}carboxamidinehydrochloride (386 mg, 1.4 mmol) were used.

m/z 501 (MH+)

Example 243 Preparation of{2-[(6-Amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-(4-phenylimidazolyl)pyrimidin-2-yl]aminedihydrochloride

Made using the same procedure as for[4-(2,4-dichlorophenyl)-5-pyrazolylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}aminedihydrochloride except that1-(2,4-dichlorophenyl)-3-(dimethylamino)-2-(4-phenylimidazolyl)prop-2-en-1-one(502 mg, 1.3 mmol) and amino{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}carboxamidine hydrochloride (386 mg, 1.4 mmol) were used andthe crude product was purified on silica gel (5% MeOH/CH₂Cl₂)

m/z 563 (MH+)

Example 244 Preparation of{2-[(6-Amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-(2,4-dimethylimidazolyl)pyrimidin-2-yl]aminedihydrochloride

Made using the same procedure as for[4-(2,4-dichlorophenyl)-5-pyrazolylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}aminedihydrochloride except that1-(2,4-dichlorophenyl)-3-(dimethylamino)-2-(2,4-dimethylimidazolyl)prop-2-en-1-one(439 mg, 1.3 mmol) andamino{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}carboxamidinehydrochloride (386 mg, 1.4 mmol) were used and the crude product waspurified by reversed-phase HPLC (gradient of 95:5 H₂O:MeCN to 5:95H₂O:MeCN).

m/z 515 (MH+)

Example 245 Preparation of Polymer-bound N-BOC-ethylenediamine

To a suspension of Merrifield resin (30 g, 21 mmol) and NMP (200 mL) wasadded 4-hydroxy-2-methoxybenzaldehyde (6.4 g, 42 mmol) and K₂CO₃ (8.7 g,63 mmol). The resulting mixture was heated at 120° C. with shaking for16 h. The resulting light brown mixture was filtered and the resin waswashed with H₂O, NMP and CH₂Cl₂. The resin was dried under vacuum at 40°C. for 12 h.

To a suspension of the resin-bound aldehyde (30 g, 21 mmol) and (MeO)₃CH(200 mL) was added N-BOC-ethylenediamine (6.7 mL, 42 mmol). Theresulting mixture was shaken at 23° C. for 12 h, filtered and washedwith CH₂Cl₂. The resin-bound imine was used immediately, slightly moistwith CH₂Cl₂.

To a suspension of the resin-bound imine (30 g, 21 mmol) andMeOH/CH₂Cl₂/HOAc (200 mL, 2:2:1) was added borane-pyridine complex (6.8mL, 67 mmol). The resulting mixture was shaken at 23° C. for 12 h,filtered and washed with NMP and CH₂Cl₂. The resin was dried undervacuum at 30° C. for 12 h to yield polymer-bound N-BOC-ethylenediamine.

Example 246 Preparation of Polymer-bound(2aminoethyl)(5-nitro(2-pyridyl))amine

To a suspension of polymer-bound N-BOC-ethylenediamine (30 g, 21 mmol),NMP (200 mL) and iPr₂NEt (18.3 mL, 105 mmol) at 23° C. was added2-chloro-5-nitropyridine (16.6 g, 105 mmol). The resulting mixture washeated at 120° C. with shaking for 12 h, filtered and washed with NMP,H₂O and CH₂Cl₂.

To the resin-bound, N-BOC-protected amine was added a solution of2,6-lutidine and CH₂Cl₂ (100 mL, 150 mmol), followed by a solution ofTMSOTf and CH₂Cl₂ (100 niL, 100 mmol). The resulting mixture was shakenat 23° C. for 3 h, filtered and washed with MeOH, Et₃N and CH₂Cl₂. Theresin was air dried to yield polymer-bound(2aminoethyl)(5-nitro(2-pyridyl))amine.

The air dried resin (10 mg) was suspended in 80% TFA/CH2Cl2 (1 mL) for 1h, filtered, washed with CH₂Cl₂ (1 mL) and concentrated under a streamof air to yield a light yellow residue.

m/z 183 (MH+)

Example 247 Preparation of Polymer-bound{2-[(5-nitro(2-pyridyl))amino]ethyl}[(2-nitrophenyl)sulfonyl]amine

To a suspension of resin-bound (2aminoethyl)(5-nitro(2-pyridyl))amine(30 g, 21 mmol), CH₂Cl₂ (250 mL) and iPr2NEt (18.3 mL, 105 mmol) at 23°C. was added 2-nitrobenzenesulfonyl chloride (23.3 g, 105 mmol). Theresulting mixture was shaken at 23° C. for 6 h, filtered, washed withNMP, H₂O and CH₂Cl₂ and air dried to yield polymer-bound{2-[(5-nitro(2-pyridyl))amino]ethyl}[(2-nitrophenyl)sulfonyl]amine.

The air dried resin (10 mg) was suspended in 80% TFA/CH₂Cl₂ (1 mL) for 1h, filtered, washed with CH₂Cl₂ (1 mL) and concentrated under a streamof air to yield a light yellow residue.

m/z 368 (MH+)

Example 248 Preparation of Polymer-bound[2-(dimethylamino)ethyl]{2-[(5-nitro(2-pyridyl))amino]ethyl}[(2-nitrophenyl)sulfonyl]amine

To a solution of Ph3P (11 g, 42 mmol) and CH₂Cl₂ (20 mL) at 23° C. wasadded DIAD (6.6 mL, 42 mmol) and the resulting yellow solution wasmaintained at 23° C. for 30 min. To this solution was added2-(dimethylamino)ethanol (4.2 mL, 42 mmol) and the resulting solutionwas maintained at 23° C. for 5 min. then added to a suspension ofresin-bound{2-[(5-nitro(2-pyridyl))amino]ethyl}[(2-nitrophenyl)sulfonyl]amine (3.0g, 2.1 mmol) and CH2Cl2 (30 mL). The resulting mixture was shaken at 23°C. for 12h, filtered, washed with NMP, H₂O and CH2Cl2 and air dried toyield polymer-bound[2-(dimethylamino)ethyl]{2-[(5-nitro(2-pyridyl))amino]ethyl}[(2-nitrophenyl)sulfonyl]amine.

The air dried resin (10 mg) was suspended in 80% TFA/CH₂Cl₂ (1 mL) for 1h, filtered, washed with CH₂Cl₂ (1 mL) and concentrated under a streamof air to yield a light yellow residue

m/z 439 (MH+)

Example 249 Preparation of Polymer-bounddimethyl[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)ethyl]amine

To a suspension of resin-bound[2-(dimethylamino)ethyl]{2-[(5-nitro(2-pyridyl))amino]ethyl}[(2-nitrophenyl)sulfonyl]amine(3.0 g, 2.1 mmol) and DMF (30 mL) at 23° C. was added H₂O (2 drops),K₂CO₃ (2.9 g, 21 mmol) and PhSH (2.2 mL, 21 mmol). The resulting mixturewas shaken at 23° C. for 12 h, filtered, washed with NMP, H₂O and CH₂Cl₂and air dried to yield polymer-bound dimethyl[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)ethyl]amine.

The air dried resin (10 mg) was suspended in 80% TFA/CH₂Cl₂ (1 mL) for 1h, filtered, washed with CH₂Cl₂ (1 mL) and concentrated under a streamof air to yield a light yellow residue.

m/z 254 (MH+)

Example 250 Preparation of Polymer-boundamino[2-(dimethylamino)ethyl]{2-[(5-nitro(2-pyridyl))amino]ethyl}carboxamidinehydrochloride

To a suspension of polymer-bound dimethyl[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)ethyl]amine (3.0 g, 2.1mmol), NMP (30 mL) and iPr₂NEt (3.7 mL, 21 mmol) at 23° C. was added1H-pyrazole-1-carboxamidine hydrochloride (3.1 g, 21 mmol). Theresulting mixture was heated at 90° C. for 18 h, filtered, washed withNMP, H₂O and CH₂Cl₂ and air dried to yield polymer-bound amino[2-(dimethylamino)ethyl]{2-[(5-nitro(2-pyridyl))amino]ethyl}carboxamidinehydrochloride.

The air dried resin (10 mg) was suspended in 80% TFA/CH₂Cl₂ (1 mL) for 1h, filtered, washed with CH₂Cl₂ (1 mL) and concentrated under a streamof air to yield a light yellow residue

m/z 296 (MH+)

Example 251 Preparation of[4-(2,4-Dichlorophenyl)-5-imidazolylpyrimidin-2-yl][2-(dimethylamino)ethyl]{2-(5-nitro(2-pyridyl))amino]ethyl}aminetrihydrochloride

To a suspension of resin-bound amino[2-(dimethylamino)ethyl]{2-[(5-nitro(2-pyridyl))amino]ethyl}carboxamidinehydrochloride (3.0 g, 2.1 mmol) and NMP (30 mL) at 23° C. was added7-methyl-1,5,7-triazabicyclo[4.4.0]dec-5-ene (1.5 mL, 10.5 mmol) and1-(2,4-dichlorophenyl)-3-(dimethylamino)-2-imidazolylprop-2-en-1-one(XVII, 1.3 g, 4.2 mmol). The resulting mixture was heated at 120° C. for20 h, filtered, washed with NMP, H₂O and CH₂Cl₂ and air dried to yieldresin-bound[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl][2-(dimethylamino)ethyl]{2-[(5-nitro(2-pyridyl))amino]ethyl}aminetrihydrochloride.

The resin was suspended in 80% TFA/CH₂Cl₂ (30 mL) and shaken at 23° C.for 1.5 h, filtered and concentrated under a stream of air. Theresulting crude material was purified by reversed-phase HPLC (gradientof 95:5 H₂O:MeCN to 5:95 H₂O:MeCN) and the recovered material wasdissolved in MeCN/0.5M HCl (3 mL, 1:1), frozen and lyophilized to yielda yellow solid.

m/z 543 (MH+)

Example 252 Preparation of Polymer-bound{2-[(5-nitro(2-pyridyl))amino]ethyl}[(2-nitrophenyl)sulfonyl](2-pyrrolidinylethyl)amine

Made using the same procedure as for polymer-bound [2-(dimethylamino)ethyl]{2-[(5-nitro(2-pyridyl))amino]ethyl}[(2-nitrophenyl)sulfonyl]amineexcept that 1-(2-hydroxy-ethyl)pyrrolidine (4.9 mL, 42 mmol) was used.

m/z 465 (MH+)

Example 253 Preparation of Polymer-bound(5-nitro(2-pyridyl)){2[(2-pyrrolidinylethyl)amino]ethyl}amine

Made using the same procedure as for polymer-bound dimethyl[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)ethyl]amine except thatresin-bound{2-[(5-nitro(2-pyridyl))amino]ethyl}[(2-nitrophenyl)sulfonyl](2-pyrrolidinylethyl)aminewas used.

m/z 280 (MH+)

Example 254 Preparation of Polymer-boundamino{2-[(5-nitro(2-pyridyl))amino]ethyl}(2-pyrrolidinylethyl)carboxamidinehydrochloride

Made using the same procedure as for polymer-bound amino[2-(dimethyl-amino)ethyl]{2-[(5-nitro(2-pyridyl))amino]ethyl}carboxamidinehydrochloride except that resin-bound(5-nitro(2-pyridyl)){2[(2-pyrrolidinylethyl)amino]ethyl}amine was used.

m/z 322 (MH+)

Example 255 Preparation of[4-(2,4-Dichlorophenyl)-5-imidazolylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}(2-pyrrolidinylethyl)aminetrihydrochloride

Made using the same procedure as for[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl][2-(dimethylamino)ethyl]{2-[(5-nitro(2-pyridyl))amino]ethyl}aminetrihydrochloride except that resin-boundamino{2-[(5-nitro(2-pyridyl))amino]ethyl}(2-pyrrolidinylethyl)carboxamidinehydrochloride was used.

m/z 569 (MH+)

Example 256 Preparation of Polymer-bound(2-morpholin-4-ylethyl){2-[(5-nitro(2-pyridyl))amino]ethyl}[(2-nitrophenyl)sulfonyl]amine

Made using the same procedure as for polymer-bound[2-(dimethylamino)ethyl]{2-[(5-nitro(2-pyridyl))amino]ethyl}[(2-nitrophenyl)sulfonyl]amineexcept that 4-(2-hydroxy-ethyl)morpholine (5.1 mL, 42 mmol) was used.

m/z 481 (MH+)

Example 257 Preparation of Polymer-bound{2-[(2-morpholin-4-ylethyl)amino]ethyl}(5-nitro(2-pyridyl))amine

Made using the same procedure as for polymer-bound dimethyl[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)ethyl]amine except thatresin-bound(2-morpholin-4-ylethyl){2-[(5-nitro(2-pyridyl))amino]ethyl}[(2-nitrophenyl)sulfonyl]aminewas used.

m/z 296 (MH+)

Example 258 Preparation of Polymer-boundamino{2-morpholin-4-ylethyl){2-[(5-nitro(2-pyridyl))amino]ethyl}carboxamidinehydrochloride

Made using the same procedure as for polymer-bound amino[2-(dimethyl-amino)ethyl]{2-[(5-nitro(2-pyridyl))amino]ethyl}carboxamidinehydrochloride except that resin-bound{2-[(2-morpholin-4-ylethyl)amino]ethyl}(5-nitro(2-pyridyl))amine wasused.

m/z 338 (MH+)

Example 259 Preparation of[4-(2,4-Dichlorophenyl)-5-imidazolylpyrimidin-2-yl](2-morpholin-4-ylethyl){2-[(5-nitro(2-pyridyl))amino]ethyl}aminetrihydrochloride

Made using the same procedure as for[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl][2-(dimethylamino)ethyl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine trihydrochloride except that resin-boundamino{2-morpholin-4-ylethyl){2-[(5-nitro(2-pyridyl))amino]ethyl}carboxamidinehydrochloride was used.

m/z 585 (MH+)

Example 260 Preparation of6-[(2-{[4-(2,4-Dichlorophenyl)-5-(5-chloro-2-oxohydropyridyl)pyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carbonitrilehydrochloride

To a solution of1-{2-(2,4-dichlorophenyl)-1-[(dimethylamino)methylene]-2-oxoethyl}-5-chlorohydropyridin-2-one(XVIII, 482 mg, 1.3 mmol) and DMF (10 mL) at 23° C. was addedamino{2-[(5-cyano(2-pyridyl))amino]ethyl}carboxamidine hydrochloride(337 mg, 1.4 mmol) followed by Cs2CO3(652 mg, 2.0 mmol) and theresulting mixture was heated at 100° C. for 16 h. The DMF was removedunder reduced pressure and the resulting residue was purified byrecrystallization (CH2Cl2/Et2O/hexanes) to yield a yellow solid whichwas dissolved in MeCN/0.5M HCl (3 mL, 1:1), frozen and lyophilized toyield a yellow solid.

m/z 513 (MH+)

Example 261 Preparation of ethyl6-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-4-phenylpyridine-3-carboxylate

1. Preparation of diethyl (2Z)-3-phenylpent-2-ene-1,5-dioate.

A solution of iodobenzene (1.08 ml, 9.67 mmol) in DMA (5 ml) was addeddropwise to a solution of diethylglutaconate (2 g, 10.74 mmol), Pd(OAc)₂(250 mg, 1.07 mmol), NaOAc (880 mg, 10.74 mmol) in DMA (5 ml) at 115° C.under argon. After heating for 8 hours at 130° C., the reaction wascooled, diluted with CH₂CI₂ (60 ml), and washed with water (4×10 ml).The organic layer was washed with sat. aq. NaHCO₃ (20 ml), brine (20ml), dried with Na₂SO₄, filtered, and concentrated under reducedpressure. The dark oil was purified by column chromatography usingCH₂Cl₂ as the eluent. The product oil was dried overnight in vacuogiving diethyl (2Z)-3-phenylpent-2-ene-1,5-dioate in 28% yield.

2. Preparation of diethyl(3E)-2-(hydroxymethylene)-3-phenylpent-3-ene-1,5-dioate.

A stirred mixture of NaH (158.4 mg, 6.6 mmol) and ethyl formate (1.07ml, 13.2 mmol) in Et₂O (5 ml) were refluxed under argon for 15 min.Diethyl (2Z)-3-phenylpent-2-ene-1,5-dioate in Et₂O (5 ml) was addeddropwise to the above solution over 5 min at rt. The reaction was heatedto reflux for 12 hours. The heterogeneous yellow mixture was dilutedwith Et₂O (100 ml), washed with sat. NH₄Cl (40 ml), half sat. NH₄Cl (40ml), brine (20 ml), dried with Na₂SO₄, filtered, and concentrated underreduced pressure. The product diethyl(3E)-2-(hydroxymethylene)-3-phenylpent-3-ene-1,5-dioate was obtained in97% yield and could be used without further purification.

3. Preparation of ethyl 6-oxo-4-phenylhydropyridine-3-carboxylate.

A mixture of diethyl(3E)-2-(hydroxymethylene)-3-phenylpent-3-ene-1,5-dioate (0.62 g, 2.13mmol) was dissolved in gla. acetic acid (1 ml), toluene (1 ml), and abs.ethanol (3 ml). Ammonium acetate (0.07 g, 9.08 mmol) and flame dried 4 Åpowder molecular sieves (0.4 g) were added to the stirred solution. Theresulting mixture was stirred for 44-46 hours at 90-95° C. under argon.After 24 hours of heating, additional reagents were added includingammonium acetate (0.07 g, 9.08 mmol), acetic acid (1 ml), and flamedried 4 Å powder molecular sieves (0.4 g). On cooling EtOAc (80 ml) wasadded with stirring for 15 minutes. The sieves were filtered and washedwith EtOAc (2×10 ml). The filtrate was concentrated under reducepressure. To the crude material was added EtOAc (100 ml). The organiclayer was then washed with distilled water (2×30 ml), sat. aq. NaHCO₃(30 ml), water (30 ml), brine (30 ml), dried with Na₂SO₄, filtered, andconcentrated under reduced pressure. The oil was purified by columnchromatography using 5% MeOH in CH₂Cl₂ as the eluent. The product oilwas dried overnight in vacuo giving ethyl6-oxo-4-phenylhydropyridine-3-carboxylate in 72% yield.

4. Preparation of ethyl 6-chloro-4-phenylpyridine-3-carboxylate.

To the dry ethyl 6-oxo-4-phenylhydropyridine-3-carboxylate (141 mg, 0.58mmol) was added phosphorous oxychloride (10 ml) followed byNN-dimethylacetamide (1 drop). The reaction mixture was stirred for 12hours at 100° C. under argon. The phosphorous oxychloride was removedunder reduced pressure. The crude product was taken up indichloromethane (2×25 ml), and the solvent was removed under vacuum. Theglass was dried in vacuo 3-4 hour giving ethyl6-chloro-4-phenylpyridine-3-carboxylate in 97% yield. The crude materialis contaminated with phosphorous residue and is used with out furtherpurification.

5. Preparation of ethyl6-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-4-phenylpyridine-3-carboxylate

The crude material above, ethyl 6-chloro-4-phenylpyridine-3-carboxylate(140 mg, 0.56 mmol) was mixed with(2-aminoethyl)(5-nitro(2-pyridyl))amine (408 mg, 2.24 mmol), Hünig'sbase (390 ul), and DMA (2 ml) for 48 hours at 70-75° C. with stirringunder argon. The reaction was followed by TLC and HPLC. When judgedcomplete, the reaction was diluted with EtOAc (100 ml) and washed withsat. aq. NaHCO₃ (5×30 ml), brine (30 ml), dried with Na₂SO₄, filtered,and concentrated under reduced pressure. The yellow solid was purifiedby column chromatography using 5% MeOH in CH₂Cl₂ as the cluent. Theproduct was dried overnight in vacuo giving ethyl6-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-4-phenylpyridine-3-carboxylatein 70% yield.

HPLC: 3.52 min (>95% purity) (HP-1 method)

MS: M+H=408.2 (C₂₁H₂₁N₅O₄+H=408)

Example 262 Preparation of[5-((1E)-1-aza-2-morpholin-4-ylprop-1-enyl)-4-(2,4-dichlorophenyl)pyrimidin-2-yl]{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amine

A. 2-[2-(2,4-dichlorophenyl)-2-oxoethyl]isoindoline-1,3-dione

1 mmol of 2,4-dichlorophenacyl chloride in DMF was added drop wise to 2mmol of phthalimide and 2 mmol of Cs₂CO₃ in DMF at room temperature forfourteen hours and then the reaction mixture was concentrated in vacuoand diluted with water and ethyl acetate. The ethyl acetate layer wasconcentrated and then purified by trituration with diethyl ether.

B.2-{2-(2,4-dichlorophenyl)-1-[(dimethylamino)methylene]-2-oxoethyl}isoindoline-1,3-dione

1 mmol of 2-[2-(2,4-dichlorophenyl)-2-oxoethyl]isoindoline-1,3-dione washeated to 80° C. in N,N-dimethylformamidedimethyl acetal for six hours.The reaction mixture was concentrated in vacuo and purified bytrituration with diethyl ether.

C.2-{N-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]carbamoyl}benzoicacid

1 mmol of2-{2-(2,4-dichlorophenyl)-1-[(dimethylamino)methylene]-2-oxoethyl}isoindoline-1,3-dione,1 mmol ofamino{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}carboxamidine, and 3mmol of Cs₂CO₃ were dissolved in DMF and heated to 90° C. for fourteenhours. The reaction mixture was concentrated in vacuo and diluted withwater and ethyl acetate. The solution was extracted three times withethyl acetate and dried over sodium sulfate.

D.2-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]isoindoline-1,3-dione

1 mmol of2-{N-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]carbamoyl}benzoicacid was heated to 120° C. in glacial acetic acid for four hours andconcentrated in vacuo.

E. [5-amino-4-(2,4-dichlorophenyl)pyrimidin-2-yl]{2-[(6-amino-

1 mmol of2-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]isoindoline-1,3-dioneand 20 mmol of hydrazine were stirred in ethanol at 75° C. for two hoursand purified by column chromatography eluting with 5-10%methanol/methylene chloride.

F.N-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]acetamide

1 mmol of[5-amino-4-(2,4-dichlorophenyl)pyrimidin-2-yl]{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amineand 1 mmol of acetic anhydride were stirred at room temperature for fourhours in THF. The reaction mixture was concentrated in vacuo and dilutedwith water and ethyl acetate. The solution was extracted three timeswith ethyl acetate, dried over sodium sulfate, and purified by columnchromatography eluting with 5-10% methanol/methylene chloride.

G.1-{[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]amino}ethane-1-thione

1 mmol ofN-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]acetamideand 2 mmol of Lawesson's reagent were stirred in 2 ml of DME at 80° C.The reaction mixture was concentrated in vacuo and diluted with waterand ethyl acetate. The solution was extracted three times with ethylacetate, dried over sodium sulfate, and purified by columnchromatography eluting with 5-10% methanol/methylene chloride.

H.[5-((1Z)-1-aza-2-morpholin-4-ylprop-1-enyl)-4-(2,4-dichlorophenyl)pyrimidin-2-yl]{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amine

1 mmol of1-{[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]amino}ethane-1-thionewas heated to 90° C. in morpholine and purified by column chromatographyeluting with 5-10% methanol/methylene chloride.

HPLC: 9.75 min. (100% purity)

MS: MH⁺=546.3

Example 263 Preparation of{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[6-(2,4-dichlorophenyl)-5-nitro(2-pyridyl)]amine

A. 2-(2,4-dichlorophenyl)-6-chloro-3-nitropyridine

1 mmol 2,6-dichloro-3-nitropyridine, 1.05 mmol of2,4-dichlorobenzeneboronic acid, and 3 mmol of Na₂CO₃, were dissolved in1.5 ml THF and 0.5 ml water and purged with nitrogen. 0.05 mmol of[1,1′-Bis(diphenylphosphino)ferrocne]dichloropalladium(II) was added toreaction and stirred at room temperature under nitrogen for fourteenhours. The reaction mixture was concentrated in vacuo and diluted withwater and ethyl acetate. The solution was extracted three times withethyl acetate, dried over sodium sulfate, and purified by columnchromatography eluting with 10% ethylacetate 90% hexanes.

B. (2-aminoethyl)(6-amino-5-nitro(2-pyridyl))amine

1 mmol of 2-amino-6-chloro-3-nitropyridine and 15 amnol of1,2-diaminoethane were stirred at reflux for fourteen hours. Thereaction mixture was concentrated in vacuo and solution of 1.5 mmol ofNaOH in water was added. This solution was extracted twice with 95%/5%methylene chloride/methanol. The aqueous was then saturated with saltand extracted twice with 95%/5% acetonitrile/methanol and then finallyextrated twice with 95%/5% ethylacetate/methanol. All organic fractionswere combined and dried over sodium sulfate.

C.{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[6-(2,4-dichlorophenyl)-5-nitro(2-pyridyl)]amine

1 mmol of 2-(2,4-dichlorophenyl)-6-chloro-3-nitropyridine was taken with2 mmol of (2-aminoethyl)(6-amino-5-nitro(2-pyridyl))amine and 3 mmol ofN,N-diisopropylethylamine in 2 ml of DMF at 80° C. for two hours. Thereaction mixture was concentrated in vacuo and diluted with water andethyl acetate. The solution was extracted three times with ethylacetate, dried over sodium sulfate, and purified by columnchromatography eluting with 5-10% methanol/methylene chloride.

HPLC: 8.698 min. (100% purity)

MS: MH⁺=464.1

Example 264 Preparation of6-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]-3-pyrrolino[3,4-b]pyridine-5,7-dione

A. 2-[2-(2,4-dichlorophenyl)-2-oxoethyl]isoindoline-1,3-dione

1 mmol of 2,4-dichlorophenacyl chloride in DMF was added drop wise to 2mmol of phthalimide and 2 mmol of Cs₂CO₃ in DMF at room temperature forfourteen hours and then the reaction mixture was concentrated in vacuoand diluted with water and ethyl acetate. The ethyl acetate layer wasconcentrated and then purified by trituration with diethyl ether.

B.2-{2-(2,4-dichlorophenyl)-1-[(dimethylamino)methylene]-2-oxoethyl}isoindoline-1,3-dione

1 mmol of 2-[2-(2,4-dichlorophenyl)-2-oxoethyl]isoindoline-1,3-dion washeated to 80° C. in neat N,N-dimethylformamidedimethyl acetal for sixhours. The reaction mixture was concentrated in vacuo and purified bytrituration with diethyl ether.

C.2-{N-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]carbamoyl}benzoicacid

1 mmol of2-{2-(2,4-dichlorophenyl)-1-[(dimethylamino)methylene]-2-oxoethyl}isoindoline-1,3-dione,1 mmol of amino{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}carboxamidine, and 3 mmol of Cs₂CO₃ were dissolved in DMFand heated to 90° C. for fourteen hours. The reaction mixture wasconcentrated in vacuo and diluted with water and ethyl acetate. Thesolution was extracted three times with ethyl acetate and dried oversodium sulfate.

D.2-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]isoindoline-1,3-dione

1 mmol of2-{N-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]carbamoyl}benzoicacid was heated to 120° C. in glacial acetic acid for four hours andthen concentrated in vacuo.

E.[5-amino-4-(2,4-dichlorophenyl)pyrimidin-2-yl]{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amine

1 mmol of2-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]isoindoline-1,3-dioneand 20 mmol of hydrazine were stirred in ethanol at 75° C. for two hoursand then purified by column chromatography eluting with 5-10%methanol/methylene chloride.

F.6-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]-3-pyrrolino[3,4-b]pyridine-5,7-dione

1 mmol of[5-amino-4-(2,4-dichlorophenyl)pyrimidin-2-yl]{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amine,and 2 mmol of furano[3,4-b]pyridine-5,7-dione were stirred at roomtemperature for four hours. 2 mmol of HBTU, and 3 mmol ofN,N-diisopropylethylamine were added to solution and left for six hoursat room temperature. The reaction mixture was concentrated in vacuo anddiluted with water and ethyl acetate. The solution was extracted threetimes with ethyl acetate, dried over sodium sulfate, and purified bycolumn chromatography eluting with 5-10% methanol/methylene chloride.

HPLC: 7.829 min. (97.32% purity)

MS: MH⁺=566.0

Example 265 Screening for GSK3 Inhibitory Activity Using a Cell-FreeAssay

Pyriridine and pyridine compounds of the present invention weredissolved in DMSO, then tested for inhibition of human GSK3β (thenucleotide sequence for human GSK3β appears in GenBank under AccessionNo. L33801). Expression of GSK3β is described, for example, in Hughes etal., Eur. J Biochem., 203:305-11 (1992), which is incorporated herein byreference.

An aliquot of 300 μl of substrate buffer (30 mM tris-HCl, 10 mM MgCl₂, 2mM DTT, 3 μg/ml GSK3β and 0.5 μM biotinylated prephosphorylatedSGSG-linked CREB peptide (Chiron Technologies PTY Ltd., Clayton,Australia) was dispensed into wells of a 96 well polypropylenemicrotiter plate. 3.5 μl /well of DMSO containing varying concentrationsof each compound to be assayed or staurosporine (a known kinaseinhibitor used as a positive control, or a negative control (i.e., DMSOonly), was added and mixed thoroughly. The reactions were then initiatedby adding 50 μl/well of 1 μM unlabeled ATP and 1-2×10⁷ cpm γ³³P-labeledATP, and the reaction was allowed to proceed for about three hours atroom temperature.

While the reaction was proceeding, streptavidin-coated Labsystems“Combiplate 8” capture plates (Labsystems, Helsinki, Finland) wereblocked by incubating them with 300 μl/well of PBS containing 1% bovineserum albumin for at least one hour at room temperature. The blockingsolution was then removed by aspiration, and the capture plates werefilled with 100 μl/well of stopping reagent (50 μM ATP/20 mM EDTA).

When the three hour enzyme reaction was finished, triplicate 100 μlaliquots of each reaction mix were transferred to three wells containingstopping solution, one well on each of the three capture plates, and thewell contents were mixed well. After one hour at room temperature, thewells of the capture plates were emptied by aspiration and washed fivetimes using PBS and a 12 channel Corning 430474 ELISA plate washer.Finally, 200 μl of Microscint-20 scintillation fluid was added to eachwell of the plate. The plates were coated with plate sealers, then lefton a shaker for 30 minutes. Each capture plate was counted in a PackardTopCount scintillation counter (Meridian, Conn.) and the results wereplotted as a function of compound concentration.

Compounds of the present invention were then screened for inhibitoryactivity against GSK3 according to this assay. The following compoundsexhibited IC₅₀s of 10 μM or less with respect to GSK3 in this cell-freeassay: (4-phenylpyrimidin-2-yl)(2-(2-pyridyl)ethyl)amine,(4-phenylpyrimidin-2-yl)[2-(2-pyridylamino)ethyl]amine,[2-(2-pyridylamino)ethyl](4-(3-pyridyl)pyrimidin-2-yl)amine,4-{2-[(2-(2-pyridyl)ethyl)amino]pyrimidin-4-yl}benzenecarbonitrile,4-{2-[(4-pyridylmethyl)amino]pyrimidin-4-yl}benzamide,4-{2-[(3-imidazol-5-ylethyl)amino]pyrimidin-4-yl}benzamide,4-(2-{[2-(2-pyridylamino)ethyl]amino}pyrimidin-4-yl)benzenecarbonitrile,4-methyl-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylicacid, 4-(2-{[(3-methylphenyl)methyl]amino}pyrimidin-4-yl)benzamide,4-(2-{[(4-aminophenyl)methyl]amino}pyrimidin-4-yl)benzamide,(5-ethyl-4-phenylpyrimidin-2-yl)[2-(2-pyridylamino) ethyl]amine,4-(2-{[(5-methylpyrazin-2-yl)methyl]aamino}pyrimidin-4-yl)benzamide,4-{2-[(3-phenoxypropyl)amino]pyrimidin-4-yl}phenol,4-{2-[(3-imidazolylpropyl) amino]pyrimidin-4-yl}benzamide,[4-(3,4-difluorophenyl)pyrimidin-2-yl][2-(2-pyridylamino)ethyl]amine,4-(2-{[(4-cyanophenyl)methyl]amino}pyrimidin-4-yl)benzamide,4-{2-[(2-phenoxyethyl)amino]pyrimidin-4-yl}benzamide,4-(2-{[(3-methoxyphenyl) methyl]amino}pyrimidin-4-yl)benzamide,4-(2-{[(4-methoxyphenyl)methyl]amino}pyrimidin-4-yl)benzamide,4-(2-{[2-(4-fluorophenyl)ethyl]amino}pyrimidin-4-yl)benzamide,[2-(2,5-dimethoxyphenyl)ethyl](4-(3-pyridyl)pyrimidin-2-yl)amine,[4-(4-nitrophenyl)pyrimidin-2-yl][2-(2-pyridylamino)ethyl]amine,{2-[(5-nitro(2-pyridyl))amino]ethyl}(4-pyrazin-2-ylpyrimidin-2-yl)amine,ethyl 4-(2-furyl)-2-[(2-(2-pyridyl)ethyl)amino]pyrimidine-5-carboxylate,4-(2-{[(3-chlorophenyl)methyl]amino}pyrimidin-4-yl)benzamide,[4-(4-chlorophenyl)-5-methylpyrimidin-2-yl][2-(2-pyridylamino)ethyl]amine,4-{2-[(3-benzimidazolylpropyl)amino]pyrimidin-4-yl}phenol,4-{2-[(4-phenylbutyl) amino]pyrimidin-4-yl}benzamide,4-(2-{[2-(3-methoxyphenyl)ethyl]amino}pyrimidin-4-yl)benzamide,4-{2-[(3-phenoxypropyl)amino]pyrimidin-4-yl}benzamide,4-(2-{[(3-nitro-phenyl)methyl]amino}pyrimidin-4-yl)benzamide,4-[2-({2-[(5-nitro-2-pyridyl)amino]ethyl}amino)pyrimidin-4-yl]phenol,3-[2-({2-[(5-nitro-2-pyridyl)amino]ethyl}amino)pyrimidin-4-yl]phenol,4-(2-{[2-(3-chlorophenyl)ethyl]amino}pyrimidin-4-yl)benzamide,4-{2-[(naphthylmethyl)amino]pyrimidin-4-yl}benzamide,[5-(4-fluoro-phenyl)pyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine,4-(2-{[3-(4-chloro-phenoxy)propyl]amino}pyrimidin-4-yl)phenol,[4-(4-imidazolylphenyl)pyrimidin-2-yl][2-(2-pyridylamino)ethyl]amine,4-(2-{[3-(2-aminobenzimidazolyl)propyl]amino}pyrimidin-4-yl)phenol,4-(2-{[2-(2,5-dimethoxyphenyl)ethyl]amino}pyrimidin-4-yl)benzene-carbonitrile,[4-(2,4-dichlorophenyl)pyrimidin-2-yl][2-(2-pyridylamino)ethyl]amine,3-[2-({2-[(5-nitro-2-pyridyl)amino]ethyl}amino)pyrimidin-4-yl]benzenecarbonitrile,4-(2-{[3-(3-methylphenoxy)propyl]amino}pyrimidin-4-yl)benzamide,4-{2-[(2-(2H-benzo[3,4-d]1,3-dioxolen-5-yl)ethyl)amino]pyrimidin-4-yl}benzamide,4-(2-{[2-(4-nitrophenyl)ethyl]amino}pyrimidin-4-yl)benzamide,4-(2-{[(2,6-dimethoxyphenyl)methyl]amino}pyrimidin-4-yl)benzamide,4-(2-{[(3,4-dimethoxyphenyl)methyl]amino}pyrimidin-4-yl)benzamide,[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]benzylamine,ethyl4-(4-fluorophenyl)-2-[(2-(2-pyridyl)ethyl)amino]pyrimidine-5-carboxylate,[4-(2,4-dimethyl(1,3-thiazol-5-yl))pyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine,[4-(4-methyl-1-phenylpyrazol-3-yl)pyrimidin-2-yl][2-(2-pyridyl-amino)ethyl]amine,4-[2-({[3-(trifluoromethyl)phenyl]methyl}amino)pyrimidin-4-yl]benzamide,4-[2-({[4-(trifluoromethyl)phenyl]methyl}amino)pyrimidin-4-yl]benzamide,4-(2-{[(3,5-dichlorophenyl)methyl]amino}pyrimidin-4-yl)benzamide,4-{2-[4-benzylpiperazinyl]pyrimidin-4-yl}benzamide,4-(2-{[(2,4-dichlorophenyl) methyl]amino}pyrimidin-4-yl)benzamide, ethyl4-(4-cyanophenyl)-2-[(2-(2-pyridyl)ethyl)amino]pyrimidine-5-carboxylate,[2-(2-pyridylamino)ethyl]{4-[4-(trifluoromethoxy)phenyl]pyrimidin-2-yl}amine,{6-[(2-methoxyethyl)amino]-5-nitro(2-pyridyl)}{2-[(5-nitro(2-pyridyl))amino]ethyl}amine,4-(2-{[3-(3-methoxyphenoxy) propyl]amino}pyrimidin-4-yl)benzamide, ethyl4-(4-methoxyphenyl)-2-[(2-(2-pyridyl)ethyl)amino]pyrimidine-5-carboxylate,[(3-methylphenyl)methyl][2-({2-[(5-nitro-(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]amine,4-(2-{[3-(4-phenylimidazolyl)propyl]amino}pyrimidin-4-yl)benzenecarbonitrile,4-{2-[(3-benzimidazolylpropyl)amino]pyrimidin-4-yl}-2-chlorophenol,4-[2-({3-[4-(2,4-dichlorophenyl)imidazolyl]propyl}amino)pyrimidin-4-yl]benzamide,4-[2-({3-[4-(3-methoxyphenyl)imidazolyl]propyl}amino)pyrimidin-4-yl]benzamide,(3-benzimidazolylpropyl)[4-(4-imidazolyl-phenyl)pyrimidin-2-yl]amine,N-{4-[2-({2-[(5-nitro-2-pyridyl)amino]ethyl}amino)pyrimidin-4-yl]phenyl}acetamide,4-(2-{[3-(4-chlorophenoxy)propyl]amino}pyrimidin-4-yl)benzamide,4-(2-{[(4-bromophenyl)methyl]amino}pyrimidin-4-yl)benzamide,4-[2-({[4-(4-fluorophenyl)phenyl]methyl}amino)pyrimidin-4-yl]benzamide,4-(2-{[(3-bromophenyl)methyl]amino}pyrimidin-4-yl)benzamide,6-{[2-({5-nitro-6-[benzylamino]-2-pyridyl}amino)ethyl]amino}pyridine-3-carbonitrile,ethyl 4-(4-cyanophenyl)-2-{[2-(pyrimidin-2-ylamino)ethyl]amino}pyrimidine-5-carboxylate,4-{2-[4-(2-methoxy-phenyl)piperazinyl]pyrimidin-4-yl}benzamide,4-(2-{[2-(benzothiazol-2-ylamino)ethyl]amino}pyrimidin-4-yl)benzamide,ethyl4-(3-nitrophenyl)-2-[(2-(2-pyridyl)ethyl)amino]pyrimidine-5-carboxylate,6-methyl-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-4-phenylpyrimidine-5-carboxylicacid, 4-{2-[(2,2-diphenylethyl)amino]pyrimidin-4-yl}benzamide,4-(2-{[(3,4,5-trimethoxyphenyl)methyl]aamino}pyrimidin-4-yl)benzamide,methyl2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-4-(3-pyridyl)pyrimidine-5-carboxylate,4-[2-({[3-(3-aminophenyl)phenyl]methyl}amino)pyrimidin-4-yl]benzamide,4-[2-({[4-(3-aminophenyl)phenyl]methyl}amino)pyrimidin-4-yl]benzamide,4-{2-[(3-(2-naphthyloxy)propyl)amino]pyrimidin-4-yl}benzamide,4-{2-[(3-(6-quinolyloxy)propyl)-amino]pyrimidin-4-yl}benzamide,[(3-chlorophenyl)methyl][2-({2-[(5-nitro(2-pyridyl))-amino]ethyl}amino)pyrimidin-4-yl]amine,[(4-chlorophenyl)methyl][2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]amine,ethyl4-(4-cyanophenyl)-2-{[2-(3-methoxyphenyl)ethyl]amino}pyrimidine-5-carboxylate,ethyl2-({2-[(5-amino(2-pyridyl))amino]ethyl}amino)-4-(4-cyanophenyl)pyrimidine-5-carboxylate,4-[5-nitro-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]benzenecarbonitrile,ethyl4-[2-({2-[(5-nitro-2-pyridyl)amino]ethyl}amino)pyrimidin-4-yl]benzoate,ethyl4-(3-nitrophenyl)-2-{[2-(2-pyridylamino)ethyl]amino}pyrimidine-5-carboxylate,N-benzyl(4-{2-[(2-(2-pyridyl)ethyl)amino]pyrimidin-4-yl}phenyl)carboxamide,{2-[(5-nitro(2-pyridyl))amino]ethyl}{5-nitro-6-[benzylamino](2-pyridyl)}amine,4-(2-methoxyphenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylicacid,4-[2-({[3-(3-methoxyphenyl)phenyl]methyl}amino)pyrimidin-4-yl]benzamide,4-[2-({2-[(5-nitro-2-pyridyl)amino]ethyl}amino)pyrimidin-4-yl]benzenesulfonamide,4-(2-{[3-(2,4-dichloro-phenoxy)propyl]amino}pyrimidin-4-yl)benzamide,4-(2-{[3-(3,4-dichlorophenoxy)propyl]amino}pyrimidin-4-yl)benzamide,4-(2-{[3-(3-phenylphenoxy) propyl]amino}pyrimidin-4-yl)benzamide,4-(2-{[(3-{3-[(methylamino)methyl]phenyl}phenyl)methyl]amino}pyrimidin-4-yl)benzamide,ethyl4-(3-fluorophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate,[4-phenyl-5-benzyl-pyrimidin-2-yl][2-(2-pyridylamino)ethyl]amine,4-(2-{[3-(3-bromophenoxy) propyl]amino}pyrimidin-4-yl)benzamide,4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-5-phenylpyrimidin-4-yl]phenol,4-(2,4-dichlorophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carbonitrile,4-(3,4-dichlorophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carbonitrile,6-[(2-{[4-(4-cyanophenyl)-5-(ethoxycarbonyl)pyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carboxylicacid, ethyl4-(3-cyanophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate,[(3,5-dichlorophenyl)methyl][2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]amine,[(2,4-dichlorophenyl)methyl][2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]amine,ethyl 4-(4-carbamoylphenyl)-6-ethyl-2-{[2-(2-pyridylamino)ethyl]amino}pyrimidine-5-carboxylate, ethyl4-(4-carbamoylphenyl)-6-ethyl-2-{[2-(pyrimidin-2-ylamino)ethyl]amino}pyrimidine-5-carboxylate,ethyl4-(3,4-dimethylphenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate,N-(2-methoxyethyl){4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}carboxamide,4-{2-[({3-[3-(acetylamino)phenyl]phenyl}methyl)amino]pyrimidin-4-yl}benzamide,ethyl 4-(4-cyanophenyl)-2-{[2-(2-quinolylamino)ethyl]amino}pyrimidine-5-carboxylate,4-[2-({[3,5-bis(trifluoromethyl)phenyl]methyl}amino)pyrimidin-4-yl]benzamide,ethyl4-(2,4-difluorophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate,(4-{[(3-bromophenyl)methyl]amino}pyrimidin-2-yl){2-[(5-nitro(2-pyridyl))amino]ethyl}amine,2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(3,4-dichlorophenyl)pyrimidine-5-carbonitrile,methyl6-[(2-{[4-(4-cyanophenyl)-5-(ethoxy-carbonyl)pyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carboxylate,4-{2-[({3-[3-(trifluoromethyl)phenyl]phenyl}methyl)amino]pyrimidin-4-yl}benzamide,[4-(4-benzimidazolylphenyl)pyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine,ethyl2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-4-(3-nitrophenyl)pyrimidine-5-carboxylate,ethyl4-naphthyl-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate,ethyl2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-4-(3-quinolyl)pyrimidine-5-carboxylate,ethyl4-(4-cyanophenyl)-2-[(2-{[5-(N-ethylcarbamoyl)(2-pyridyl)]amino}ethyl)amino]pyrimidine-5-carboxylate,benzyl{[4-(2-{[2-(2-pyridylamino)ethyl]amino}pyrimidin-4-yl)phenyl]sulfonyl}amine, ethyl4-(4-cyanophenyl)-2-[(2-{[6-(methylamino)-5-nitro(2-pyridyl)]amino}ethyl)amino]pyrimidine-5-carboxylate,{4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}-N-(oxolan-2-ylmethyl)carboxamide,N-(1-carbamoyl-2-phenylethyl)[4-methyl-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-5-yl]carboxamide,N-(1-carbamoyl-2-phenyl-ethyl)(4-{2-[(2-(2-pyridyl)ethyl)amino]pyrimidin-4-yl}phenyl)carboxamide,ethyl4-(3,4-dimethoxyphenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate,{4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}-N-benzylcarboxamide,{4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}-N-(3-pyridylmethyl)carboxamide,{4-[4-(4,5-dichloroimidazol-2-yl)phenyl]pyrimidin-2-yl}{2-[(5-nitro(2-pyridyl))amino]ethyl}amine,ethyl4-(4-butoxyphenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate,ethyl4-{[(2-chlorophenyl)methyl]amino}-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate,6-(2-fluorophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-4-phenyl-pyrimidine-5-carboxylicacid,{4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}-N-(2-thienylmethyl)carboxamide,ethyl2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-4-[3-(trifluoromethyl)phenyl]pyrimidine-5-carboxylate,ethyl4-(3,4-dichlorophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate,ethyl 4-(3,5-dichlorophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate,{4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}-N-(4-piperidylmethyl)carboxamide,(6-{[(2,4-dichlorophenyl)methyl]amino}-5-nitro(2-pyridyl)){2-[(5-nitro(2-pyridyl))amino]ethyl}amine,ethyl4-(4-carbamoylphenyl)-6-ethyl-2-({2-[(3-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate,ethyl4-[4-(diethylamino)phenyl]-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate,{4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}-N-(2-phenylethyl)carboxamide,N-[(3-methylphenyl)methyl]{4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}carboxamide,2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4,6-trichlorophenyl)pyrimidine-5-carboxylicacid, ethyl2-({2-[(5-nitro(2-pyridyl))amino]ethyl}aamino)-4-(4-phenylphenyl)pyrimidine-5-carboxylate,{4-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}-N-benzylcarboxamide,N-[(5-methylpyrazin-2-yl)methyl]{4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}carboxamide,N-[(3-fluorophenyl)methyl]{4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}carboxamide,ethyl2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-4-(4-sulfamoyl-phenyl)pyrimidine-5-carboxylate,N-[(4-fluorophenyl)methyl]{4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}carboxamide,[4-(2-{[(3-bromo-phenyl)methyl]amino}pyrimidin-4-yl)phenyl]-N-[(3-methylphenyl)methyl]carboxamide,ethyl4-(5-bromo(3-pyridyl))-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate,N-(3-imidazolylpropyl){4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}carboxamide,tert-butyl6-[(2-{[4-(4-cyanophenyl)-5-(ethoxycarbonyl)pyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carboxylate,N-[(3-bromophenyl)methyl](4-{2-[(3-imidazolylpropyl)amino]pyrimidin-4-yl}phenyl)carboxamide,ethyl 4-[(2,4-dichlorophenyl)amino]-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate, ethyl4-(4-carbamoylphenyl)-6-ethyl-2-({3-[(5-nitro(2-pyridyl))amino]propyl}amino)pyrimidine-5-carboxylate,{4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}-N-(2-phenylcyclopropyl)carboxamide,N-[(4-methoxyphenyl)methyl]{4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}carboxamide,2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-6-(4-nitrophenyl)-4-phenylpyrimidine-5-carboxylicacid,2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-6-(4-nitrophenyl)-4-(4-pyridyl)pyrimidine-5-carboxylicacid, ethyl4-(4-carbamoylphenyl)-6-ethyl-2-[(2-{[5-(trifluoromethyl)(2-pyridyl)]amino}ethyl)amino]pyrimidine-5-carboxylate,ethyl4-(4-cyanophenyl)-2-[(2-{[5-(morpholin-4-ylcarbonyl)(2-pyridyl)]amino}ethyl)amino]pyrimidine-5-carboxylate,N-[(3-chlorophenyl)methyl]{4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}carboxamide,N-[(3,4-difluorophenyl)methyl]{4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}carboxamide, ethyl4-[4-(4-methylpiperazinyl)phenyl]-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate,4-cyclohexyl-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-6-(4-nitrophenyl)pyrimidine-5-carboxylicacid,{4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}-N-[(3-nitrophenyl)methyl]carboxamide,ethyl4-{[(3-bromophenyl)methyl]amino}-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate,N-[(3-bromophenyl)methyl][4-(2-{[2-(3-methoxyphenyl)ethyl]amino}pyrimidin-4-yl)phenyl]carboxamide,N-(naphthyl-methyl){4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}carboxamide,4-(3-cyanophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-6-(4-nitrophenyl)pyrimidine-5-carboxylicacid,N-[(3,4-dimethoxyphenyl)methyl]{4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}carboxamide,N-[(2,3-dimethoxyphenyl)methyl]{4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}carboxamide,4-(4-methoxyphenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-6-(4-nitrophenyl)pyrimidine-5-carboxylicacid,N-[(3-bromophenyl)methyl]{4-[2-({2-[(6-methoxy(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}carboxamide,{4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}-N-{[3-(trifluoromethyl)phenyl]methyl}carboxamide,N-[(3,5-dichlorophenyl)methyl]{4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}carboxamide,N-[(3,4-dichlorophenyl)methyl]{4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}carboxamide,ethyl4-(4-cyanophenyl)-2-{[2-({5-nitro-6-[benzylamino](2-pyridyl)}amino)ethyl]amino}pyrimidine-5-carboxylate,ethyl4-[3,5-bis(trifluoromethyl)phenyl]-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate,4,6-bis(4-nitrophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylicacid,[4-(2-{[2-(2,5-dimethoxyphenyl)ethyl]amino}pyrimidin-4-yl)phenyl]-N-[(3-bromophenyl)methyl]carboxamide,2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-4-(3-nitrophenyl)-6-(4-nitrophenyl)pyrimidine-5-carboxylicacid,N-[(3-bromophenyl)methyl]{4-[2-({2-[(4-nitrophenyl)amino]ethyl}amino)pyrimidin-4-yl]phenyl}carboxamide,N-[(3-bromophenyl)methyl]{4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}carboxamide,N-[(4-bromophenyl)methyl]{4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}carboxamide,{4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}-N-[(4-sulfamoylphenyl)methyl]carboxamide,N-[2-(2,4-dichlorophenyl)ethyl]{4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}carboxamide,N-[(3-bromophenyl)methyl][4-(2-{[2-(2-quinolylamino)ethyl]amino}pyrimidin-4-yl)phenyl]carboxamide,2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-6-(4-nitrophenyl)-4-(4-quinolyl)pyrimidine-5-carboxylicacid,N-(2,2-diphenylethyl){4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}carboxamide,N-[(3-bromophenyl)methyl]{4-[2-({3-[(5-nitro(2-pyridyl))amino]propyl}amino)pyrimidin-4-yl]phenyl}carboxamide,{4-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}aamino)pyrimidin-4-yl]phenyl}-N-[(3-bromophenyl)methyl]carboxamide,2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-6-(4-nitrophenyl)-4-[4-(trifluoromethyl)phenyl]pyrimidine-5-carboxylicacid,N-[(3-bromophenyl)methyl](4-{2-[(2-{[5-(trifluoromethyl)(2-pyridyl)]amino}ethyl)amino]pyrimidin-4-yl}phenyl)carboxamide,[(3-bromophenyl)methyl]({4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}sulfonyl)amine, andN-[(3-iodophenyl)methyl]{4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}carboxamide,1-[2-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]aamino}-4-(2,4-dichlorophenyl)-5-pyrimidinyl]-2-piperazinone,1-[2-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-4-(2,4-dichlorophenyl)-5-pyrimidinyl]-4-ethyl-3-methyl-2-piperazinone,1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-2-(2,4-dichlorophenyl)-3-pyridinyl]-4-methyl-2-piperazinone,1-[2-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-4-(2,4-dichlorophenyl)-5-pyrimidinyl]-4-methyl-2-piperazinone,6-[(2-{[6-(2,4-dichlorophenyl)-5-(4-methyl-2-oxo-1-piperazinyl)-2-pyridinyl]amino}ethyl)amino]nicotinonitrile,1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-2-(4-ethylphenyl)-3-pyridinyl]-4-methyl-2-piperazinone,6-[(2-{[6-(4-ethylphenyl)-5-(4-methyl-2-oxo-1-piperazinyl)-2-pyridinyl]amino}ethyl)amino]nicotinonitrile,6-({2-[[6-(4-ethylphenyl)-5-(4-methyl-2-oxo-1-piperazinyl)-2-pyridinyl](methyl)amino]ethyl}amino)nicotinonitrile,6-[(2-{[4-(2,4-dichlorophenyl)-5-(4-methyl-2-oxo-1-piperazinyl)-2-pyrimidinyl]amino}ethyl)amino]nicotinonitrile,1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-2-(2,4-difluorophenyl)-3-pyridinyl]-4-methyl-2-piperazinone,4-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]aamino}-3-(4-methyl-2-oxo-1-piperazinyl)-2-pyridinyl]benzonitrile,1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-2-(2,4-dichlorophenyl)-3-pyridinyl]-4-isopropyl-2-piperazinone,1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-2-(2,4-dichlorophenyl)-3-pyridinyl]-4-ethyl-2-piperazinone,1-{6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-2-[2-(trifluoromethyl)phenyl]-3-pyridinyl-}4-methyl-2-piperazinone,1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-2-(2-bromophenyl)-3-pyridinyl]-4-methyl-2-piperazinone,1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)propyl]amino}-2-(2,4-dichlorophenyl)-3-pyridinyl]-4-methyl-2-piperazinone,1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)propyl]amino}-2-(2,4-dichlorophenyl)-3-pyridinyl]-4-methyl-2-piperazinone,1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)-1-methylethyl]amino}-2-(2,4-dichlorophenyl)-3-pyridinyl]-4-methyl-2-piperazinone,1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)-1-methylethyl]amino}-2-(2,4-dichlorophenyl)-3-pyridinyl]-4-methyl-2-piperazinone,1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)-1,1-dimethylethyl]amino}-2-(2,4-dichloro-phenyl)-3-pyridinyl]-4-methyl-2-piperazinone,1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl][2-(1-pyrrolidinyl)ethyl]amino}-2-(2,4-dichlorophenyl)-3-pyridinyl]-4-methyl-2-piperazinone,1-[6-[3-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)propyl]-2-(2,4-dichlorophenyl)-3-pyridinyl-4-methyl-2-piperazinone,1-[6-[[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl](methyl)amino]-2-(2,4-dichlorophenyl)-3-pyridinyl]-4-methyl-2-piperazinone,1-[6-[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethoxy]-2-(2,4-dichlorophenyl)-3-pyridinyl]-4-methyl-2-piperazinone,1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}oxy)ethyl]amino}-2-(2,4-dichlorophenyl)-3-pyridinyl]-4-methyl-2-piperazinone,1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl][2-(4-morpholinyl)ethyl]amino}-2-(2,4-dichlorophenyl)-3-pyridinyl]-4-methyl-2-piperazinone,1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-2-(2,4-dichlorophenyl)-3-pyridinyl]-2-piperazinone,1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-2-(2,4-dichlorophenyl)-3-pyridinyl]-4-cyclopropyl-2-piperazinone,1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-2-(2,4-dichlorophenyl)-3-pyridinyl]-4-cyclohexyl-2-piperazinone,1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}aamino)ethyl]amino}-2-(4-bromophenyl)-3-pyridinyl]-4-methyl-2-piperazinone,1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-2-(2,4-dichlorophenyl)-3-pyridinyl]-3,4-dimethyl-2-piperazinone,1-(2-(2,4-dichlorophenyl)-6-{[2-({5-[hydroxy(oxido)amino]-6-methoxy-2-pyridinyl}amino)ethyl]amino}-3-pyridinyl)-4-methyl-2-piperazinone,1-(2-(2,4-dichlorophenyl)-6-{[2-({5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-3-pyridinyl)-4-methyl-2-piperazinone,4-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-3-(4-methyl-2-oxo-1-piperazinyl)-2-pyridinyl]benzamide,2-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-2-(2,4-dichlorophenyl)-3-pyridinyl]hexahydropyrrolo[1,2-a]pyrazin-3(4H)-one,1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-2-(2,4-dichlorophenyl)-3-pyridinyl]-4-(methylsulfonyl)-2-piperazinone,4-acetyl-1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-2-(2,4-dichlorophenyl)-3-pyridinyl]-2-piperazinone,2-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-2-(2,4-dichlorophenyl)-3-pyridinyl]hexahydropyrrolo[1,2-a]pyrazin-3(4H)-one,2-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-2-(2,4-dichlorophenyl)-3-pyridinyl]hexahydropyrrolo[1,2-a]pyrazin-3(4H)-one,2-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-2-(2,4-dichlorophenyl)-3-pyridinyl]hexahydropyrrolo[1,2-a]pyrazin-1(2H)-one,2-[6-{[2-({6-amino-5-[hydroxy(oxido) amino]-2-pyridinyl}amino)ethyl]amino}-2-(2,4-dichlorophenyl)-3-pyridinyl]hexahydropyrrolo-[1,2-a]pyrazin-1(2H)-one,1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-2-(2,4-difluorophenyl)-3-pyridinyl]-4-ethyl-2-piperazinone,4-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-3-(4-ethyl-2-oxo-1-piperazinyl)-2-pyridinyl]benzonitrile,1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)propyl]amino}-2-(2,4-dichlorophenyl)-3-pyridinyl]-4-ethyl-2-piperazinone,1-(6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-2-phenyl-3-pyridinyl)-4-methyl-2-piperazinone,1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-2-(4-chlorophenyl)-3-pyridinyl]-4-methyl-2-piperazinone,3-amino-i-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-2-(2,4-dichlorophenyl)-3-pyridinyl]-2-piperidinone,1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-2-(3-chlorophenyl)-3-pyridinyl]-4-methyl-2-piperazinone,1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-2-(2,4-dichlorophenoxy)-3-pyridinyl]-4-methyl-2-piperazinone,1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-2-(2,4-dichlorophenyl)-3-pyridinyl]-4-(2,2,2-trifluoroethyl)-2-piperazinone,4-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-2-(2,4-dichloro-phenyl)-3-pyridinyl]-3-morpholinone,1-{6-[2-(6-Amino-5-nitro-pyridin-2-ylamino)ethylamino]-[2,3′]bipyridinyl-3-yl}-4-methyl-piperazin-2-one,1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-2-(2,4-dichlorophenyl)-3-pyridinyl]-2-piperidinone,1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-2-(4-chloro-2-methylphenyl)-3-pyridinyl]-4-methyl-2-piperazinone,1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-2-(4-methoxyphenyl)-3-pyridinyl]-4-methyl-2-piperazinone,and1-[6-{[2-({6-amino-5-[hydroxy(oxido)amino]-2-pyridinyl}amino)ethyl]amino}-2-(3-furyl)-3-pyridinyl]-4-methyl-2-piperazinone.

The following compounds exhibited IC₅₀s of 1 μM or less with respect toGSK3 in this cell-free assay:4-{2-[(2-(2-pyridyl)ethyl)amino]pyrimidin-4-yl}benzamide,4-{2-[(2-phenylpropyl)amino]pyrimidin-4-yl}benzamide,4-(2-{[2-(2-pyridylamino) ethyl]amino}pyrimidin-4-yl)benzamide,4-(2-{[2-(pyrimidin-2-ylamino)ethyl]amino}pyrimidin-4-yl)benzamide,(5-nitro-4-phenylpyrimidin-2-yl)[2-(2-pyridylamino)ethyl]amine,4-(2-{[3-(4-nitroimidazolyl)propyl]amino}pyrimidin-4-yl)phenol,4-(2-{[2-(2-methoxyphenyl)ethyl]amino}pyrimidin-4-yl)benzamide,4-[2-({2-[(5-cyano-2-pyridyl)amino]ethyl}amino)pyrimidin-4-yl]benzamide,2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-4-phenylpyrimidine-5-carbonitrile,4-[2-({2-[(6-methoxy-2-pyridyl)amino]ethyl}amino)pyrimidin-4-yl]benzamide,4-(2-{[3-(4,5-dichloroimidazolyl)propyl]amino}pyrimidin-4-yl)phenol,4-(2-{[3-(4-nitroimidazolyl)propyl]amino}pyrimidin-4-yl)benzamide,4-{2-[(3-benzimidazolylpropyl)amino]pyrimidin-4-yl}benzamide,4-[2-({2-[(4-amino-5-cyano-pyrimidin-2-yl)amino]ethyl}amino)pyrimidin-4-yl]benzamide,4-{2-[(3-benzimidazolylpropyl)amino]pyrimidin-4-yl}-2-methoxyphenol,4-(2-{[2-(2,5-dimethoxyphenyl)ethyl]amino}pyrimidin-4-yl)benzamide,4-(2-{[2-(2,3-dimethoxy-phenyl)ethyl]amino}pyrimidin-4-yl)benzamide,4-(2-{[3-(4-methoxyphenoxy) propyl]amino}pyrimidin-4-yl)benzamide,4-[2-({2-[(5-nitro-2-pyridyl)amino]ethyl}amino)pyrimidin-4-yl]benzamide,{2-[(5-nitro(2-pyridyl))amino]ethyl}(5-nitro-4-phenyl-pyrimidin-2-yl)amine,4-(2-{[2-(2-quinolylamino) ethyl]amino}pyrimidin-4-yl)benzamide,4-(4-cyanophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carbonitrile,4-(2-{[3-(4,5-dichloroimidazolyl)propyl]amino}pyrimidin-4-yl)benzamide,4-{2-[(2-{[5-(aminothioxomethyl)-2-pyridyl]amino}ethyl)amino]pyrimidin-4-yl}benzamide,4-[2-({3-[(5-nitro-2-pyridyl)amino]propyl}amino)pyrimidin-4-yl]benzamide,4-(2-{[3-(4-phenylimidazolyl)propyl]amino}pyrimidin-4-yl)benzamide,4-{2-[(3-naphthyloxypropyl)amino]pyrimidin-4-yl}benzamide,4-(2-{[3-(5,6-dimethylbenzimidazolyl)propyl]amino}pyrimidin-4-yl)benzamide,[4-(4-imidazolylphenyl)pyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine,4-{2-[(2-{[5-(trifluoromethyl)-2-pyridyl]amino}ethyl)amino]pyrimidin-4-yl}benzamide,4-(4-cyanophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxamide,4-(4-cyanophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylicacid,2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}aamino)-4-(2,4-dichlorophenyl)pyrimidine-5-carboxylicacid, ethyl2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-4-(4-pyridyl)pyrimidine-5-carboxylate,ethyl2-({2-[(5-cyano(2-pyridyl))amino]ethyl}amino)-4-(4-cyanophenyl)pyrimidine-5-carboxylate,4-[2-({3-[3-(trifluoromethyl)phenoxy]propyl}amino)pyrimidin-4-yl]benzamide,[4-(4-cyanophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-5-yl]-N-methylcarboxamide,methyl4-(4-cyanophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate,[4-(4-morpholin-4-ylphenyl)pyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine,ethyl4-(4-methylphenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate,{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}{5-nitro-6-[benzylamino](2-pyridyl)}amine,2-({2-[(5-nitro-(2-pyridyl))amino]ethyl}amino)-4-(4-nitrophenyl)pyrimidine-5-carboxylicacid, ethyl4-(4-fluorophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate,4-[5-imidazolyl-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]benzene-carbonitrile,4-[5-imidazol-2-yl-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]benzenecarbonitrile,ethyl2-({2-[(4-amino-5-cyanopyrimidin-2-yl)amino]ethyl}amino)-4-(4-cyanophenyl)pyrimidine-5-carboxylate,[4-(2,4-dimethyl-phenyl)-5-imidazolylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine,2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidine-5-carbonitrile,ethyl4-(4-cyanophenyl)-2-({2-[(4-nitrophenyl)amino]ethyl}amino)pyrimidine-5-carboxylate,[4-(4-cyanophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-5-yl]-N,N-dimethylcarboxamide,ethyl4-(4-cyanophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate,[4-(2,4-dichlorophenyl)-5-ethylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine,ethyl 4-(4-ethylphenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate, ethyl4-(4-methoxyphenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate,4-[5-(methylsulfonyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]benzenecarbonitrile,4-(2-{[3-(5,6-dichlorobenzimidazolyl)propyl]amino}pyrimidin-4-yl)benzamide,4-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-5-imidazolyl-pyrimidin-4-yl]benzenecarbonitrile,4-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-5-imidazol-2-ylpyrimidin-4-yl]benzenecarbonitrile,N-(cyanomethyl)[4-(4-cyanophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-5-yl]carboxamide,4-[5-(3-methyl(1 ,2,4-oxadiazol-5-yl))-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]benzenecarbonitrile, ethyl4-(4-chlorophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate,ethyl4-(3,4-difluoro-phenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate,N-(2-aminoethyl)[4-(4-cyanophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-5-yl]carboxamide,ethyl4-(4-cyanophenyl)-2-({2-[(4-methyl-5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate,ethyl2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyll}amino)-4-(4-cyanophenyl)pyrimidine-5-carboxylate,[4-(4-cyanophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-5-yl]-N-(2-hydroxyethyl)carboxamide, 2-hydroxyethyl4-(4-cyanophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate,ethyl2-({2-[(4-amino-5-nitropyrimidin-2-yl)amino]ethyl}amino)-4-(4-cyanophenyl)pyrimidine-5-carboxylate,ethyl4-[4-(methylethyl)phenyl]-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate,ethyl4-[4-(dimethylamino)phenyl]-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate,ethyl4-(2H-benzo[3,4-d]1,3-dioxolen-5-yl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate,ethyl2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-4-(4-nitrophenyl)pyrimidine-5-carboxylate,ethyl4-(4-methylthiophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate,ethyl4-(4-cyanophenyl)-2-[(2-{[5-(trifluoromethyl)(2-pyridyl)]amino}ethyl)amino]pyrimidine-5-carboxylate,ethyl 4-(2-naphthyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate,N-butyl[4-(4-cyanophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-5-yl]carboxamide,N-(tert-butyl)[4-(4-cyano-phenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-5-yl]carboxamide,ethyl4-(4-carbamoylphenyl)-2-({2-[(5-cyano(2-pyridyl))amino]ethyl}amino)-6-ethyl-pyrimidine-5-carboxylate,tert-butyl 4-(4-cyanophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate,N-(carbamoylmethyl)[4-(4-cyanophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-5-yl]carboxamide,ethyl4-(4-cyano-phenyl)-6-ethyl-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate,butyl4-(4-cyanophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate,ethyl 4-{[(4-cyanophenyl)methyl]amino}-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate, ethyl4-{[(3-cyanophenyl)methyl]amino}-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate,ethyl4-[4-(tert-butyl)phenyl]-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate,4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-5-[benzylamino]pyrimidin-4-yl]benzene-carbonitrile,[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine,[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine,4-(4-cyanophenyl)-6-(3-fiuryl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylicacid, 4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-5-(piperazinylcarbonyl)pyrimidin-4-yl]benzenecarbonitrile,ethyl4-(4-imidazolylphenyl)-2-({2-[((5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate,4-[5-(morpholin-4-ylcarbonyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]benzenecarbonitrile,ethyl4-(4-(2-furyl)phenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate,ethyl 2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-4-(4-(1,3-oxazol-5-yl)phenyl)pyrimidine-5-carboxylate, ethyl2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-4-(4-(1,2,4-triazol-4-yl)phenyl)pyrimidine-5-carboxylate,N-[2-(dimethylamino)ethyl][4-(4-cyanophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-5-yl]carboxamide,2-(dimethylamino) ethyl4-(4-cyano-phenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate, ethyl2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-4-[4-(trifluoromethyl)phenyl]pyrimidine-5-carboxylate,ethyl4-(2,4-dichlorophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate,ethyl4-(4-(1H-1,2,3,4-tetraazol-5-yl)phenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate,ethyl4-(4-carbamoylphenyl)-6-ethyl-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate,4-(4-cyanophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-6-phenylpyrimidine-5-carboxylicacid,{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl]amine,{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amine,ethyl4-(4-bromophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate,ethyl4-[4-(methyl-sulfonyl)phenyl]-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate,ethyl2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(4-(1,3-oxazol-5-yl)phenyl)pyrimidine-5-carboxylate,N-[2-(dimethylamino)ethyl][4-(4-cyanophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-5-yl]-N-methylcarboxamide,N-(1-carbamoyl-2-hydroxyethyl)[4-(4-cyanophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-5-yl]carboxamide, 3-(dimethylamino)propyl4-(4-cyanophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate,ethyl2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidine-5-carboxylate,2-(dimethylamino)ethyl2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(4-cyanophenyl)pyrimidine-5-carboxylate,[2-(dimethyl-amino)ethoxy]-N-[4-(4-cyanophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-5-yl]carboxamide,ethyl2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-4-[4-(trifluoromethoxy)phenyl]pyrimidine-5-carboxylate,ethyl4-(4-morpholin-4-ylphenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate,[4-(4-cyano-phenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-5-yl]-N-benzyl-carboxamide,ethyl 4-(6-morpholin-4-yl(3-pyridyl))-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino) pyrimidine-5-carboxylate,[4-(4-cyanophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-5-yl]-N-(4-pyridylmethyl)carboxamide,phenylmethyl4-(4-cyanophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate,4-(4-cyanophenyl)-6-(4-fluorophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylicacid,N-[(3-methoxyphenyl)methyl]{4-[2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}carboxamide,4-[5-{3-[2-(dimethylamino)ethyl](1,2,4-oxadiazol-5-yl)}-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]benzenecarbonitrile,N-[(3-bromophenyl)methyl][4-(2-{[2-(pyrimidin-2-ylamino)ethyl]amino}pyrimidin-4-yl)phenyl]carboxamide,4-(dimethyl-amino)butyl4-(4-cyanophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate,4,6-bis(4-cyanophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylicacid, ethyl 2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(4-morpholin-4-ylphenyl)pyrimidine-5-carboxylate,4-(3-hydroxyphenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-6-(4-nitrophenyl)pyrimidine-5-carboxylicacid, 2-morpholin-4-ylethyl4-(4-cyanophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate,4-(4-cyanophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-6-(4-nitrophenyl)pyrimidine-5-carboxylicacid,4-(4-cyanophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-6-(3-nitrophenyl)pyrimidine-5-carboxylicacid,N-[(3-bromophenyl)methyl]{4-[2-({2-[(5-cyano(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]phenyl}carboxamide,ethyl4-(4-carbamoylphenyl)-2-({2-[(5-nitro-(2-pyridyl))amino]ethyl})-6-(4-pyridyl)pyrimidine-5-carboxylate,2-(dimethylamino)ethyl2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichloro-phenyl)pyrimidine-5-carboxylate,2-[bis(2-hydroxyethyl)amino]ethyl4-(4-cyanophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate,{4-[2-({2-[(4-amino-5-cyanopyrimidin-2-yl)amino]ethyl}amino)pyrimidin-4-yl]phenyl}-N-[(3-bromo-phenyl)methyl]carboxamide,4-(4-carboxyphenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-6-(4-nitrophenyl)pyrimidine-5-carboxylic acid,2-hydroxy-3-morpholin-4-yl-propyl4-(4-cyanophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidine-5-carboxylate,ethyl4-(4-cyanophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-6-(4-nitrophenyl)pyrimidine-5-carboxylate,(2-{5-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl](1,2,4-oxadiazol-3-yl)}ethyl)dimethylamine, ethyl4-(4-carbamoylphenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-6-(4-nitrophenyl)pyrimidine-5-carboxylate,[4-(4-imidazolylphenyl)pyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine,4-[5-imidazolyl-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-4-yl]benzenecarbonitrile,4-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-5-imidazolylpyrimidin-4-yl]benzenecarbonitrile,[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine,4-[2-({2-[(5-nitro-2-pyridyl)amino]ethyl}amino)-7a-hydro-1,2,4-triazolo[1,5-a]pyrimidin-7-yl]benzenecarbonitrile,{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichloro-phenyl)-5-imidazolylpyrimidin-2-yl]amine,[4-(2,4-dichlorophenyl)-5-imidazol-2-yl-pyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine,6-[(2-{[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carbonitrile,[5-benzotriazolyl-4-(2,4-dichlorophenyl)pyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine,[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]methan-1-ol,[4-(2,4-dichlorophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)-pyrimidin-5-yl]methan-1-ol,2-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]isoindoline-1,3-dione,[5-amino-4-(2,4-dichlorophenyl)pyrimidin-2-yl]{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amine,{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-morpholin-4-yl-pyrimidin-2-yl]amine,{2-[(6-amino-5-nitro(2-pyridyl))aamino]ethyl}{4-(2,4-dichloro-phenyl)-5-[5-(trifluoromethyl)(1,2,3,4-tetraazolyl)]pyrimidin-2-yl}amine,1-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]pyrrolidine-2,5-dione,[4-(2,4-dichlorophenyl)-5-pyrazolylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine,[4-(2,4-dichlorophenyl)-5-(4-methylimidazolyl)pyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}aamine,[4-(2,4-dichlorophenyl)-5-(2,4-dimethylimidazolyl)pyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine,6-[(2-{[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carbonitrile,{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-(morpholin-4-ylmethyl)pyrimidin-2-yl]amine,{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-piperazinylpyrimidin-2-yl]amine,{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(4-ethylphenyl)-5-imidazolylpyrimidin-2-yl]amine,1-[4-(2,4-dichlorophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-5-yl]hydropyridin-2-one,[5-benzimidazolyl-4-(2,4-dichlorophenyl)pyrimidin-2-yl]{2-[(5-nitro-(2-pyridyl))amino]ethyl}amine,{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl]methylamine,{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-(4-pyridyl)pyrimidin-2-yl]amine,{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-(4-methylpiperazinyl)pyrimidin-2-yl]amine,[4-(2,4-dichlorophenyl)-5-(2-methylimidazolyl)pyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine,{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-(2-methylimidazolyl)pyrimidin-2-yl]amine,{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-(4-phenylimidazolyl)pyrimidin-2-yl]amine,{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-(2,4-dimethylimidazolyl)pyrimidin-2-yl]amine,[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl](2-{[5-(trifluoromethyl)(2-pyridyl)]amino}ethyl)amine,[4-(2,4-dichlorophenyl)-5-piperazinylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine,[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl][2-(dimethylamino)ethyl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine,1-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]-4-methylpiperazine-2,6-dione,[4-(2,4-dichlorophenyl)-5-(1-methylimidazol-2-yl)pyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine,1-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichloro-phenyl)pyrimidin-5-yl]-3-morpholin-4-ylpyrrolidine-2,5-dione,1-[4-(2,4-dichlorophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-5-yl]-4-methylpiperazine-2,6-dione,1-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]-3-(dimethylamino)pyrrolidine-2,5-dione,{5-imidazol-2-yl-4-[4-(trifluoromethyl)phenyl]pyrimidin-2-yl}{2-[(5-nitro(2-pyridyl))amino]ethyl}amine,{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-(1-methylimidazol-2-yl)pyrimidin-2-yl]amine,[4-(2,4-dichlorophenyl)-5-(4-methylpiperazinyl)pyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine,[4-(2,4-dichlorophenyl)-5-(morpholin-4-ylmethyl)pyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine,[4-(2,4-dichlorophenyl)-5-(4-methylimidazol-2-yl)pyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine,{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2,4-dichlorophenyl)-5-(4-methylimidazol-2-yl)pyrimidin-2-yl]amine,{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2-chlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amine,[4-(2-chloro-4-fluorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine,[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}(2-pyrrolidinylethyl)amine,[4-(2,4-dichlorophenyl)-5-imidazolylpyrimidin-2-yl](2-morpholin-4-ylethyl){2-[(5-nitro(2-pyridyl))amino]ethyl}amine,6-[(2-{[4-(2,4-dichlorophenyl)-5-(4-methylimidazol-2-yl)pyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carbonitrile,{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2-chloro-4-fluorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amine,[4-(4-ethylphenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine,[5-((1E)-1-aza-2-morpholin-4-ylprop-1-enyl)-4-(2,4-dichlorophenyl)pyrimidin-2-yl]{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amine,N-[4-(2,4-dichlorophenyl)-2-({2-[(5-nitro(2-pyridyl))amino]ethyl}amino)pyrimidin-5-yl]acetamide,[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(6-methoxy-5-nitro(2-pyridyl))amino]ethyl}amine,6-[(2-{[4-(2,4-dichlorophenyl)-5-imidazolyl-pyrimidin-2-yl]methylamino}ethyl)amino]pyridine-3-carbonitrile,6-[(2-{[4-(2,4-dichloro-phenyl)-5-imidazol-2-ylpyrimidin-2-yl]methylamino}ethyl)amino]pyridine-3-carbonitrile,[4-(2,4-dichlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]methyl{2-[(5-nitro(2-pyridyl))amino]ethyl}amine,6-[(2-{[4-(2-chloro-4-fluorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carbonitrile,[4-(4-chlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine,{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(4-chloro-2-methylphenyl)-5-imidazol-2-ylpyrimidin-2-yl]amine,{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(4-bromo-2-chlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amine,6-[(2-{[4-(4-bromo-2-chlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carbonitrile,6-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]-3-pyrrolino[3,4-b]pyridine-5,7-dione,N-[2-({2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}amino)-4-(2,4-dichlorophenyl)pyrimidin-5-yl]-2-(methylamino)acetamide,{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(4-bromo-2-chlorophenyl)-5-(4-methylimidazol-2-yl)pyrimidin-2-yl]amine,6-[(2{[4-(4-bromo-2-chlorophenyl)-5-(4-methylimidazol-2-yl)pyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carbonitrile,{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[4-(2-chloro-4-fluorophenyl)-5-(4-methylimidazol-2-yl)pyrimidin-2-yl]amine,6-[(2-{[4-(2,4-dichlorophenyl)-5-(5-chloro-2-oxohydropyridyl)pyrimidin-2-yl]amino}ethyl)amino]pyridine-3-carbonitrile,[6-(2,4-dichlorophenyl)-5-imidazolyl(2-pyridyl)]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine,{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[6-(2,4-dichlorophenyl)-5-imidazolyl(2-pyridyl)]amine,6-[(2-{[6-(2,4-dichlorophenyl)-5-imidazolyl-2-pyridyl]amino}ethyl)amino]pyridine-3-carbonitrile,{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[6-(2,4-dichlorophenyl)-5-nitro(2-pyridyl)]amine,{2-[(6-amino-5-nitro(2-pyridyl))amino]ethyl}[6-(2,4-dichlorophenyl)-5-(4-methylimidazolyl)(2-pyridyl)]amine,6-[(2-{[6-(2,4-dichlorophenyl)-5-(4-methylimidazolyl)-2-pyridyl]amino}ethyl)amino]pyridine-3-carbonitrile,and[4-(4-bromo-2-chlorophenyl)-5-imidazol-2-ylpyrimidin-2-yl]{2-[(5-nitro(2-pyridyl))amino]ethyl}amine.

Accordingly, these results demonstrate that compounds of the presentinvention exhibit inhibitory activity against GSK3.

Example 266 Screening for GSK3 Inhibitory Activity Using a Cell-BasedGlycogen Synthase Assay

CHO-HIRC cells are maintained in 10 cm tissue culture plates in Ham'sF12 medium/10% dialysed fetal bovine serum. Cells from a confluent 10 cmplate are harvested and divided into the 6 wells of a 6-well tissueculture plate to a final volume of 2 ml, of medium. The cells are leftto grow at 37° C. for 24 hours. The cells are then washed three times inHam's F12 medium containing no fetal bovine serum, and finally the cellsare left for a further 24 hours at 37° C. in 2 ml of the serum-freemedium.

At the end of this time, 20 μl of compound dissolved in DMSO is added toeach well and incubated at 37° C. After 20 minutes the medium is removedand the cells are washed once in PBS at room temperature and thenrapidly frozen in the plates in liquid nitrogen. Cells are then thawedon ice in the presence of 140 μl of lysis buffer (50 mM Tris pH 7.8; 1mM EDTA, 100 mM NaF, 25 pg/ml leupeptin, 1 mM DTT, 1 mM PMSF) per well.Cells are scraped from the plates and frozen in Eppendorf tubes on dryice. Lysates are then thawed and refrozen on dry ice.

After rethawing, lysates are spun at 14,000 g for 15 minutes. Thesupernatants are then removed and stored on ice. Each supernatant (45μl) is added to 45 μl of reaction buffer (65 mM Tris pH 7.8; 26 mM EDTA,32.5 mM KF, 9.3 mM UDP-glucose; 11 mg/ml glycogen; 500 nCi/ml¹⁴C-UDP-glucose) and a further 45 μl is added to 45 μl reactionbuffer/20 mM glucose-6-phosphate. Reactions are incubated at 30° C. for30 minutes and then spotted onto a 2 cm square 31ET chromatograph paper(Whatman). Filter papers are washed twice for 20 minutes in 66% ethanol,rinsed briefly in acetone and dried for 1 hour at room temperature.

Filters are added to 5 ml of liquid scintillant and counted in a liquidscintillation counter. The percentage of the total glycogen synthasethat is active in any lysate is expressed as 100×(cpm minusglucose-6-phosphate)/(cpm plus glucose-6-phosphate). Such values aredetermined in duplicate for 5 different concentrations of compound andfor DMSO alone, and the values are then plotted against the logarithm ofthe concentration. The concentration of compound which stimulatesglycogen synthase activity to 50% of the maximal level is determined byfitting a sigmoidal curve to the plotted data. The maximal level isdefined as that level to which glycogen synthase activity tendsasymtotically as the concentration of test compound increasessubstantially beyond the EC₅₀.

Example 267 Screening for Inhibition of Tau Protein Phosphorylation

A. Transient Transfection of COS Cells with GSK3 Expression Plasmid andTau Expression

Plasmid Construction

COS cells are maintained in T25 tissue culture flasks in high glucoseMEM medium/5% fetal bovine serum. Cells from a confluent T25 flask areharvested and 80,000 cells/well are seeded into Coming 6-well tissueculture plates in a fmal volume of 2 ml/well of medium. The cells areleft to grow at 37° C. for 48 hours. The cells are then washed twice inOpti-MEM containing no fetal bovine serum, and finally the cells areleft in 1 ml of Opti-MEM.

Polynucleotide encoding tau protein is subcloned into plasmid pSG5 underan early SV40 promoter to generate a tau expression plasmid. The cloningof cDNA encoding tau protein is generally described in Goedert et al.,EMBO Journal, 8(2):393-399 (1989), which is incorporated herein byreference. A GSK3 expression plasmid is prepared by subdloningpolynucleotide encoding GSK3β into pCG, which is an ApEVRF derivativedescribed in Giese et al., Genes & Development, 9:995-1008 (1995) andMatthias et al., Nucleic Acid Research, 17:6418 (1989), both of whichare incorporated herein by reference.

The following solutions are prepared in 1.5 ml Eppendorf tubes: SolutionA: for each transfection, 2 μg of DNA (tau expression plasmid) and 0.7μg of DNA (GSK3 expression plasmid) are diluted into 100 μl of Opti-MEM(Gibco BRL); Solution B: for each transfection, 8 μl of Lipofectaminereagent is diluted into 100 μl of Opti-MEM. The two solutions arecombined, mixed gently, and incubated at room temperature for 45 minutesto allow DNA-liposome complexes to form. For each transfection, 0.8 mlof Opti-MEM is added to the tube containing the complexes. The dilutedsolution is mixed gently and overlaid onto the rinsed cells. The cellsare incubated with the complexed DNA/Lipofectamine for 6 hours at 37° C.in a CO₂ incubator. Following incubation, 1 ml of growth medium (highglucose MEM) with 20% FBS is added to each well and incubated at 37° C.overnight. The medium is replaced with fresh, complete medium at 18hours following the start of transfection, and the cells are left togrow at 37° C. for another 48 hours.

B. Tau Phosphorylation Inhibition Assay

Two hours before harvesting, 2 μl of test compound (GSK3 inhibitor)dissolved in DMSO is added to each well and incubated at 37° C. After 2hours the medium is removed and the cells are rapidly frozen on theplates on dry ice and stored at −70° C. Cells are thawed on ice in thepresence of 200 μl of lysing buffer (1% Triton® X-100, 20 mM Tris pH7.5, 137 mM NaCl, 15% glycerol, 25 μg/ml leupeptin, 1 μg ml pepstatin-A,1 μM PMSF, 21 μg/ml aprotinin, 50 mM NaF, 50 mM β-glycerophosphate, 15mM sodium pyrophosphate, 1 mM sodium orthovanadate). The contents ofeach well are centrifuged at 14,000 g, 4° C. for 5 minutes and thesupernatants transferred to clean tubes. At this point the lysates maybe stored at −20° C.

C. ELISA to Detect Phosphorylated tau in Cell Lysates

Immulon 4 strips (Dynatech) are coated with monoclonalanti-phosphorylated tau (AT8, Polymedco, Inc.) at 5 μg/ml in PBScontaining Ca++ and Mg++, 100 μl/well. After overnight incubation at 4°C., the strips are washed twice with washing buffer (PBS containing0.05% Tween® 20) and blocked with PBS containing 1% BSA, 5% normal mouseserum and 0.05% Tween® 20 at room temperature for 1 hour. The strips arewashed 5 times with washing buffer. Lysate (100 μl) diluted 1:10 in PBScontaining 1% BSA, 0.1% NaN₃ is added into each well and incubated atroom temperature for 1 hour. After washing, 100 μl of 0.5 μg/mlbiotinylated monoclonal anti-(non-phosphorylated) tau (HT7, Polymedco,Inc.) in PBS-BSA is added into each well. Strips are washed 5 times andHRP-conjugated streptavidin is added, incubated at room temperature for30 minutes and washed extensively with washing buffer. TMB substrate(Pierce) is used for color development and the reaction is stopped byadding an equal volume of 0.8 M sulfuric acid. Strips are read on anELISA plate reader using a 450 nm filter. The concentration of compoundthat inhibits tau phosphorylation to 50% of the maximal level (i.e.,IC₅₀) is determined by fitting a sigmoidal curve to the plotted data.

Example 268 Testing the Potential of GSK3 Inhibitors to Protect PrimaryHippocampal Cells from Glutamate Excitotoxicity

Hippocampi were dissected from embryonic day 18-19 rats. The tissue wascollected in Hibernate TM media (Gibco BRL) and minced intoapproximately 1 mm pieces. Tissue was dissociated using the PapainDissociation System (Worthington Biochemical Corporation). Followingisolation the cells were resuspended in serum-free media composed ofNeurobasal TM (Gibco BRL), 2% B27 supplement (GibcoBRL), L-glutamine andantibiotics. Cells were plated in 35 mm tissue culture dishes coatedwith poly-L-lysine at a concentration of 7.5×104 cells per dish.Following 10-14 days at 37° C. in 5% CO2 cells were rinsed and fed withfresh media. The next day representative compounds of the invention wereadded to the culture media to a final concentration of between 1 nM and100 μM. Four to eight hours following compound addition the conditionedmedia was removed from cells and stored at 37° C. Cultures were rinsedtwice with HEPES-buffered balanced salt solution (HBSS) containing 10 μMglycine. Grabb and Choi, J. Neuroscience 19:1657-62 (1999). Cultureswere then exposed for 5 min at room temperature to 200 μM glutamic acidin the same HBSS. Following exposure, cultures were rinsed three timeswith the buffer and then returned to their original conditioned mediacontaining the compounds. Twenty to twenty-four hours following glutamicacid exposure, cultures were rinsed in HBSS and exposed for 10 min toTrypan Blue. This dye is taken up by dead cells. The cultures wererinsed and then fixed for 30 min in 4% paraformaldehyde . The number oflive and dead (blue nuclei) large neurons are counted (at least 200cells from each culture) by phase contrast microscopy and photographed.Using this method, compounds of this invention have been shown to becapable of significantly reducing the potential of glutamate to induceneuronal cell death.

Example 269 Evaluation of Efficacy in Diabetic Rodents The glucoseTolerance Test

Compound Formulation for Oral Dosing:

Test compounds were typically formulated for oral gavage as solutions inwater or suspensions in 1% carboxymethylcellulose/0.1% tween-80 (bothfrom Sigma Chem., MO) the day prior to administration. Some earlycompounds were formulated as solutions in 15% Captisol (a modifiedcyclodexytrin by CyDex Co., IL) following procedures common to thosebelow. For water solutions, dry and lyophilized test compound powder issolubilized in distilled water and mixed well by vortexing andsonicating. If necessary, test solution is pH adjusted with 1 N NaOH or1 N HCl and is finally sterile filtered through a syringe appended witha 0.2 micron cellulose acetate membrane (Millipore Co., MA). For oralsuspensions, the test compound powder is mixed with a fresh suspensionof 1% carboxymethylcellulose/0.1% tween-80 and extensively sonicated, pHadjusted if necessary as described above, and vortexed until particlesize is homogeneous and <10 micron in size.

Diabetic Mouse Glucose Tolerance Test:

Obese db/db (female C57BlKs/J) mice were obtained from Jackson Labs (BarHarbor, Me.) at 8 weeks of age and used for efficacy testing 1-2 weekslater. On the morning of a test, food was removed early in the morning(7-8 hrs prior to the glucose bolus). Local anesthetic (EMLA creme,Astra Pharm., MA) was applied to the end of the tail and 50-100 μl bloodsamples were obtained from snips of the tail tip and collected intoeppendorf tubes containing 5 μl 500 U/ml sodium heparin (Elkins-Sinn,NJ) with subsequent isolation of plasma. Samples were obtained atvarious intervals throughout the day for a total of 6-8 time points.Mice were randomized into treatment groups and administered the firstoral dose of test compound (0.2 ml volume) 4.5 hr prior to the glucoseand again 0.5 hr prior to administration of 0.2 ml 50% dextrose (AbbottLab., IL) via oral gavage (oGTT) or intraperitoneal injection. After thefinal blood sample about 2 hr following the glucose administration, foodwas returned to the animals.

Regulation of Basal Glycemia and Insulinemia:

Test compounds were typically orally administered to db/db mice (seeabove) or ZDF rats (Genetic Models, Inc.; Indianapolis, Ind.) in thecontext of a multi-day, multi-dose regimen or as a single bolus. The ZDFrats were received at 8 weeks of age and used for efficacy testing 1-2weeks later. Food was removed about 30 min prior to dosing and a singlebolus of test compound (dosing volume ranging from 1-8 mg/ml) wasadministered. Blood was sampled as described above at 1-6 time pointsover the next 2-3 hr. Food was returned to the animal cages followingthe blood sampling.

Primary Endpoints:

Glucose and insulin levels are measured from plasma and/or bloodsamples. Glucose levels are measured from whole blood by the One-Touchglucometer (Lifescan Co., CA) and from plasma by Beckman glucoseanalyzer. Glucose results typically reflect blood values for mouse andplasma values for rat studies. Measurement of insulin levels has beenvia ELISA (Crystal Chem. Co., IL) following the supplier's protocol.

Results Quantitation:

Efficacy may be expressed as mg/dL glucose or ng/ml insulin orrepresented as area under the curve (AUC) for plasma glucose (takenabove the normoglycemic baseline of 100 mg/dL) and insulin (taken abovethe normoinsulinemic baseline of 1 ng/mL). Typically, when expressed asAUC, the results are actually represented as reduced AUC ([(vehiclecontrol AUC—test group AUC)/vehicle control AUCX100]). Such expressionprovides a single quantitative expression of the magnitude of improvedglucose disposal and/or reduced basal hyperglycemia or insulinconservation relative to the placebo control group.

Results:

Representative compounds of the invention exhibited good in vitropotency, and when formulated in captisol and administered s.c. to mice(30 mg/kg), exhibited high bioavailability and tissue penetrance invivo. A significant reduction in basal hyperglycemia just prior to theglucose tolerance test, and significantly improved glucose disposalfollowing glucose challenge were observed. A 45-50% reduction in the AUCrelative to the control group was observed if the glucose response isquantitated by determining the area under the blood glucose curve (AUC)from −60 min to +120 min. This is comparable to the efficacy obtainedwith Troglitazone (when dosed orally for at least several days at either60 or 100 mg/kg/day). Also of significance was the observation thatinsulin levels in treated animals remained lower than in control mice.

While preferred embodiments of the invention has been illustrated anddescribed, it will be appreciated that various changes can be madetherein without departing from the spirit and scope of the invention.

1. A compound having the structure:

wherein: A is selected from the group consisting of pyridyl,pyrimidinyl, thiazolyl, indolyl, imidazolyl, oxadiazolyl, tetrazolyl,pyrazinyl, triazolyl, thiophenyl, furanyl, quinolinyl, purinyl,naphthyl, benzothiazolyl, and benzimidazolyl, which may be optionallysubstituted with from 0 to 3 substitution groups independently selectedfrom the group consisting of nitro, amino, cyano, halo, thioamido,amidino, oxamidino, alkoxyamidino, imidino, guanidino, sulfonamido,carboxyl, formyl, loweralkyl, haloloweralkyl, loweralkoxy,haloloweralkoxy, loweralkoxyalkyl, loweralkylaminoloweralkoxy,loweralkylcarbonyl, loweraralkylcarbonyl, lowerheteroaralkylcarbonyl,alkylthia, aminoalkyl and cyanoalkyl; R₁, R₂, R₃, and R₄ areindependently selected from the group consisting of hydrogen, hydroxyl,and loweralkyl, cycloloweralkyl, cyclicaniinoalkyl, alkylaminoalkyl,loweralkoxy, amino, alkylamino, alkylcarbonyl, arylcarbonyl,aralkylcarbonyl, heteroarylcarbonyl, heteroaralkylcarbonyl, aryl andheteroaryl, which may be optionally substituted with from 0 to 3substitution groups independently selected from the group consisting ofhydroxyl, nitro, amino, imino, cyano, halo, thio, thioamido, amidino,imidino, oxo, oxamidino, methoxamidino, imidino, guanidino, sulfonamido,carboxyl, formyl, loweralkyl, haloloweralkyl, loweralkoxy,haloloweralkoxy, loweralkoxyalkyl, alkylcarbonyl, arylcarbonyl,aralkylcarbonyl, heteroarylcarbonyl, heteroaralkylcarbonyl, alkylthio,aminoalkyl, and cyanoalkyl; R₅ and R₇ are independently selected fromthe group consisting of hydrogen, halo, loweralkyl, cycloalkyl, alkoxy,aminoalkoxy, alkylamino, aralkylamino, heteroaralkylamino, arylamino,heteroarylamino cycloimido, heterocycloimido, amidino, cycloamidino,heterocycloamidino, guanidinyl, aryl, biaryl, heteroaryl, heterobiaryl,heterocycloalkyl, and arylsulfonamido, which may be optionallysubstituted with from 0 to 5 substitution groups independently selectedfrom the group consisting of hydroxyl, nitro, amino, imino, cyano, halo,thio, thioamido, amidino, imidino, oxo, oxamidino, methoxamidino,imidino, guanidino, sulfonamido, carboxyl, formyl, loweralkyl,haloloweralkyl, loweralkoxy, haloloweralkoxy, loweralkoxyalkyl,atkylcarbonyl, arylcarbonyl, aralkylcarbonyl, heteroarylcarbonyl,heteroaralkylcarbonyl, alkylthio, aminoalkyl, and cyanoalkyl; and R₆ isselected from the group consisting of pyridyl, pyrimidinyl, piperazinyl,thiazolyl, indolyl, imidazolyl, oxadiazolyl, tetrazolyl, pyrazinyl,triazolyl, thienyl, furanyl, quinolinyl, pyrrolylpyridyl,benzothiazolyl, benzotriazolyl, and benzimidazolyl, which may heoptionally substituted with from 0 to 3 substitution groupsindependently selected from the group consisting of hydroxyl, nitro,amino, imino, cyano, halo, thio, thioamido, amidino, imidino, oxo,oxamidino, methoxamidino, imidino, guanidino, sulfonamido, carboxyl,formyl, loweralkyl, haloloweralkyl, loweralkoxy, haloloweralkoxy,loweralkoxyalkyl, alkylcarbonyl, arylcarbonyl, aralkylcarbonyl,heteroarylcarbonyl, heteroaralkylcarbonyl, alkylthio, aminoalkyl, andcyanoalkyl; or a pharmaceutically acceptable salt thereof.
 2. A compoundof claim 1 wherein A has the formula:

wherein R₈ and R₉ are independently selected from the group consistingof hydrogen, hydroxy, nitro, amino, cyano, halo, thioamido, amidino,oxamidino, alkoxyamidino, imidino, guanidinyl, sulfonamido, carboxyl,formyl, loweralkyl, aminoloweralkyl, loweralkylaminoloweralkyl,haloloweralkyl, loweralkoxy, haloloweralkoxy, loweralkoxyalkyl,loweralkylaminoloweralkoxy, loweralkylcarbonyl, alkylthio, aryl and,aralkyl.
 3. A compound of claim 1, wherein A is selected from the groupconsisting of aminopyridyl, nitropyridyl, aminonitropyridyl,cyanopyridyl, cyanothiazolyl, aminocyanopyridyl, trifluoromethylpyridyl,methoxypyridyl, methoxynitropyridyl, methoxycyanopyridyl andnitrothiazolyl.
 4. A compound of claim 1, wherein at least one of R₁,R₂, R₃, and R₄ is loweralkyl selected from the group consisting ofunsubstituted loweralkyl, haloloweralkyl, heterocycloaminoalkyl, andloweralkylaminoloweralkyl.
 5. A compound of claim 4, wherein at leastone of R₁, R₂, R₃, and R₄ is loweralkylaminoloweralkyl.
 6. A compound ofclaim 4, wherein R₁, R₂, and R₃ are hydrogen and R₄ is selected from thegroup consisting of methyl, ethyl, aminoethyl, dimethylaminoethyl,pyridylethyl, piperidinyl, pyrrolidinylethyl, piperazinylethyl andmorpholinylethyl.
 7. A compound of claim 1, wherein at least one of R₅and R₇ is selected from the group consisting of substituted andunsubstituted aryl, heteroaryl and biaryl.
 8. A compound of claim 7,wherein at least one of R₅ and R₇ is a moiety of the formula:

wherein R₁₀, R₁₁, R₁₂, R₁₃, and R₁₄ are independently selected from thegroup consisting of hydrogen, nitro, amino, cyano, halo, thioamido,carboxyl, hydroxy, loweralkyl, loweralkoxy, loweralkoxyalkyl,haloloweralkyl, haloloweralkoxy, aminoalkyl, alkylamino,aminoalkylalkynyl, alkylaminoalkylalkynyl, alkylthio,alkylcarbonylamino, aralkylcarbonylamino, heteroaralkylcarbonylamino,arylcarbonylamino, heteroarylcarbonylamino aminocarbonyl,loweralkylaminocarbonyl, aminoaralkyl, loweralkylaminoalkyl, aryl,heteroaryl, cycloheteroalkyl, aralkyl, alkylcarbonyloxy,arylcarbonyloxy, aralkylcarbonyloxy, arylcarbonyloxyalkyl,alkylcarbonyloxyalkyl, heteroarylcarbonyloxyalkyl,aralkycarbonyloxyalkyl, and heteroaralkcarbonyloxyalkyl, which may beoptionally substituted with from 0 to 3 substitution groupsindependently selected from the group consisting of hydroxyl, nitro,amino, imino, cyano, halo, thio, thioamido, amidino, imidino, oxo,oxamidino, methoxamidino, imidino, guanidino, sulfonamido, carboxyl,formyl, loweralkyl, haloloweralkyl, loweralkoxy, haloloweralkoxy,loweralkoxyalkyl, alkylcarbonyl, arylcarbonyl, aralkylcarbonyl,heteroarylcarbonyl, heteroaralkylcarbonyl, alkylthio, aminoalkyl, andcyanoalkyl.
 9. A compound of claim 8 wherein R₁₀, R₁₁, R₁₃, and R₁₄ arehydrogen and R₁₂ is selected from the group consisting of halo,loweralkyl, hydroxy, loweralkoxy, haloloweralkyl, aminocarbonyl,alkylaminocarbonyl and cyano.
 10. A compound of claim 8 wherein R₁₁,R₁₃, and R₁₄ are hydrogen and R₁₀ and R₁₂ are independently selectedfrom the group consisting of halo, loweralkyl, hydroxy, loweralkoxy,haloloweralkyl and cyano.
 11. A compound of claim 8 wherein R₁₀, R₁₁,R₁₃, and R₁₄ are hydrogen and R₁₂ is heteroaryl.
 12. A compound of claim8 wherein R₁₀, R₁₁, R₁₃, and R₁₄ are hydrogen and R₁₂ is aheterocycloalkyl.
 13. A compound of claim 8 wherein at least one of R₁₀,R₁₁, R₁₂, R₁₃, and R₁₄ are halo and the remainder of R₁₀, R₁₁, R₁₂, R₁₃,and R₁₄ are hydrogen.
 14. A compound of claim 8 wherein at least one ofR₅and R₇ is selected from the group consisting of dichlorophenyl,difluorophenyl, trifluoromethylphenyl, chloro fluorophenyl,bromochlorophenyl, ethylphenyl, methylchlorophenyl, imidazolylphenyl,cyanophenyl, morphlinophenyl and cyanochiorophenyl.
 15. A compound ofclaim 1, wherein R₆ is a monoketopiperazinyl group having the structure:

wherein R₁₅ and R₁₆ are independently selected from the group consistingof hydrogen, loweralkyl, loweralkynyl, aryl, heteroaryl, arylloweralkyl,loweralkylarylloweralkyl, haloloweralkyl, haloarylloweralkyl carbocyclicand heterocyclic; or R₁₆ can be taken with another R₁₆ or with R₁₅ toform a carbocyclic, heterocyclic or aryl ring; and o is an integerbetween 1 and
 6. 16. A compound of claim 15, wherein R₁₅ is loweralkyl.17. A compound of claim 16, wherein R₁₅ is selected from the groupconsisting of methyl, ethyl, n-propyl, isopropyl, cyclopropyl, n-butyl,isobutyl and t-butyl.
 18. A compound of claim 15, wherein R₁₅ is takenwith R₁₆ to form a group having the structure:


19. A compound of claim 15, wherein R₁₅ is taken with R₁₆ to form agroup having the structure:


20. A compound having the structure:

wherein: R₁, R₂, R₃ and R₄ are independently selected from the groupconsisting of hydrogen, hydroxyl, loweralkyl, cyclicaminoalkyl,alkylaminoallcyl, loweralkoxy, amino, alkylamino, alkylcarbonyl,arylcarbonyl, aralkylcarbonyl, heteroarylcarbonyl,heteroaralkylcarbonyl, aryl and heteroaryl, which may be optionallysubstituted with from 0 to 3 substitution groups independently selectedfrom the group consisting of hydroxyl, nitro, amino, imino, cyano, halo,thio, thioamido, amidino, imidino, oxo, oxamidino, methoxamidino,imidino, guanidino, sulfonamido, carboxyl, formyl, loweralkyl,haloloweralkyl, loweralkoxy, haloloweralkoxy, loweralkoxyalkyl,alkylcarbonyl, arylcarbonyl, aralkylcarbonyl, heteroarylcarbonyl,heteroaralkylcarbonyl, alkylthio, aminoalkyl, and cyanoalkyl; R₅ isselected from the group consisting of hydrogen, halo, loweralkyl,cycloalkyl, alkoxy, aminoalkoxy, alkylamino, aralkylamino,heteroaralkylamino, arylamino, heteroarylamino cycloimido,heterocycloimido, amidino, cycloamidino, heterocycloamidino, guanidinyl,aryl, biaryl, heteroaryl, heterobiaryl, heterocycloalkyl, andarylsulfonamido, which may be optionally substituted with from 0 to 5substitution groups independently selected from the group consisting ofhydroxyl, nitro, amino, imino, cyano, halo, thio, thioamido, amidino,imidino, oxo, oxamidino, methoxamidino, imidino, guanidino, sulfonamido,carboxyl, formyl, loweralkyl, haloloweralkyl, loweralkoxy,haloloweralkoxy, loweralkoxyalkyl, alkylcarbonyl, arylcarbonyl, aralkylearbonyl, heteroarylcarbonyl, heteroaralkylcarbonyl, alkylthio,aminoalkyl, and cyanoalky; R₆ is selected from the group consisting ofpyridyl, pyrimidinyl, pyrrolidinyl, thiazolyl, indolyl, imidazolyl,oxadi azolyl, tetrazolyl, pyrazinyl, triazolyl, thienyl, furanyl,quinolinyl, pyrrolylpyridyl, benzothiazolyl, benzotriazolyl, andbenzimidazolyl, which may be optionally substituted with from 0 to 3substitution groups independently selected from the group consisting ofhydroxyl, nitro, amino, imino, cyano, halo, thio, thioamido, amidino,imidino, oxo, oxamidino, methoxamidino, imidino, guanidino, sulfonamido,carboxyl, formyl, loweralkyl, haloloweralkyl, loweralkoxy,haloloweralkoxy, loweralkoxyalkyl, alkylcarbonyl, arylcarbonyl,aralkylcarbonyl, heteroarylcarbonyl, heteroaralkylcarbonyl, alkylthio,aminoalkyl, and cyanoalkyl; R₈ and R₉ are independently selected fromthe group consisting of hydrogen, hydroxy, nitro, amino, cyano, halo,thioamido, amidino, oxamidino, alkoxyamidino, imidino, guanidinyl,sulfonamido, carboxyl, formyl, loweralkyl, aminoloweralkyl,loweralkylaminoloweralkyl, haloloweralkyl, loweralkoxy, haloloweralkoxy,loweralkoxyalkyl, loweralkylaminoloweralkoxy loweralkylcarbonyl,loweraralkylcarbonyl, lowerheteroaralkylcarbonyl, alkylthio, aryl and,aralkyl; and R₁₀, R₁₁, R₁₂, R₁₃, and R₁₄ are independently selected fromthe group consisting of hydrogen, nitro, amino, cyano, halo, thioamido,carboxyl, hydroxy, loweralkyl, loweralkoxy, loweralkoxyalkyl,haloloweralkyl, haloloweralkoxy, aminoalkyl, alkylamino, alkylthio,alkylcarbonylamino, aralkylcarbonyl amino, heteroaralkylcarbonylamino,arylcarbonylamino, heteroarylcarbonylamino aminocarbonyl,loweralkylaminocarbonyl, aminoaralkyl, loweralkylaminoalkyl, aryl,heteroaryl, cycloheteroalkyl, aralkyl, and alkylcarbonyloxy,arylcarbonyloxy, aralkylcarbonyloxy, arylcarbonyloxyalkyl,alkyloxycarbonylalkyl, heteroarylcarbonyloxyalkyl,aralkycarbonyloxyalkyl, heteroaralkylcarbonyloxyalkyl, which may beoptionally substituted with from 0 to 3 substitution groupsindependently selected from the group consisting of hydroxyl, nitro,amino, imino, cyano, halo, thio, thioamido, amidino, imidino, oxo,oxamidino, methoxamidino, imidino, guanidino, sulfonamido, carboxyl,formyl, loweralkyl, haloloweralkyl, loweralkoxy, haloloweralkoxy,loweralkoxyalkyl, alkylcarbonyl, arylcarbonyl, aralkylcarbonyl,heteroarylcarbonyl, heteroaralkylcarbonyl, alkylthio, aminoalkyl, andcyanoalkyl; wherein heteroaryl is selected from the group consisting ofpyridyl, pyrimidinyl, thiazolyl, indolyl, imidazolyl, oxadiazolyl,tetrazolyl, pyrazinyl, triazolyl, thienyl, furanyl, quinolinyl,pyrrolyopyridyl, benzothiazolyl, benzotriazolyl, and benzimidazolyl,which may be optionally substituted with from 0 to 3 substitution groupsindependently selected from the group consisting of hydroxyl, nitro,amino, imino, cyano, halo, thio, thioamido, amidino, imidino, oxo,oxamidino, methoxamidino, imidino, guanidino, sulfonamido, carboxyl,formyl, loweralkyl, haloloweralkyl, loweralkoxy, haloloweralkoxy,loweralkoxyalkyl, alkylcarbonyl, arylcarbonyl, aralkylcarbonyl,heteroarylcarbonyl, heteroaralkylcarbonyl, alkylthio, aminoalkyl, andcyanoalkyl; and wherein heterocyclo is a cycloalkyl substituent havingfrom 3 to 8 ring atoms comprising from 1 to 5 heteroatoms selected fromthe group consisting of N, O and S; or a pharmaceutically acceptablesalt thereof.
 21. A compound of claim 20, wherein at least one of R₈ andR₉ is selected from the group consisting of nitro, amino, cyano,trifluoromethyl and loweralkoxy.
 22. A compound of claim 20, wherein atleast one of R₁, R₂, R₃ and R₄ is loweralkyl selected from the groupconsisting of hydrogen, unsubstituted loweralkyl, haloloweralkyl,heterocycloaminoalkyl, aminoloweralkyl and loweralkylaminoloweralkyl.23. A compound of claim 20, wherein at least one of R₁, R₂, R₃ and R₄ isloweralkylaminoloweralkyl.
 24. A compound of claim 20, wherein R₁, R₂,and R₃ are hydrogen and R₄ is selected from the group consisting ofmethyl, ethyl, aminoethyl, dimethylaminoethyl, pyridylethyl,piperidinyl, pyrrolidinylethyl, piperazinylethyl and morpholinylethyl.25. A compound of claim 20, wherein R₅ is selected from the groupconsisting of hydrogen and substituted and unsubstituted aryl,heteroaryl and biaryl.
 26. A compound of claim 20 wherein R₁₀, R₁₁, R₁₃,and R₁₄ are hydrogen and R₁₂ is selected from the group consisting ofhalo, loweralkyl, hydroxy, loweralkoxy, halolowcralkyl, aminocarbonyl,alkylaminocarbonyl and cyano.
 27. A compound of claim 20 wherein R₁₁,R₁₃, and R₁₄ are hydrogen and R₁₀ and R₁₂ are independently selectedfrom the group consisting of halo, loweralkyl, hydroxy, loweralkoxy,haloloweralkyl, aminocarbonyl and cyano.
 28. A compound of claim 20wherein R₁₀ is hydrogen or halo, R₁₁, R₁₃, and R₁₄ are hydrogen and R₁₂is heteroaryl.
 29. A compound of claim 20 wherein R₁₀ is hydrogen orhalo, R₁₁, R₁₃, and R₁₄ are hydrogen and R₁₂ is a heterocycloalkyl. 30.A compound of claim 20 wherein at least one of R₁₀, R₁₁, R₁₂, R₁₃, andR₁₄ are halo and the remainder of R₁₀, R₁₁, R₁₂, R₁₃, and R₁₄ arehydrogen.
 31. A compound of claim 20 wherein R₁₀, R₁₁, R₁₂, R₁₃, andR₁₄, taken together with the phenyl ring of structure IV, form a moietyselected from the group consisting of dieblorophenyl, difluorophenyl,trifluoromethyiphenyl, chlorofluoropheflyl, bromochlorophenyl,ethylphenyl, methylchiorophenyl, imidazolylphenyl, cyanophenyl,morpholinophenyl and cyanochlorophenyl.
 32. A compound of claim 20,wherein R₆ is a monoketopiperazinyl group having the structure:

wherein R₁₅ and R₁₆ are independently selected from the group consistingof hydrogen, loweralkyl, loweralkynyl, aryl, heteroaryl, arylloweralkyl,loweralkylarylloweralkyl, haloloweralkyl, haloarylloweralkyl carbocyclicand heterocyclic; or R₁₆ can be taken with another R₁₆ or with R₁₅ toform a carbocyclic, heterocyclic or aryl ring; and o is an integerbetween 1 and
 6. 33. A compound of claim 32, wherein R₁₅ is loweralkyl.34. A compound of claim 33, wherein R₁₅ is selected from the groupconsisting of methyl, ethyl, n-propyl, isopropyl, cyclopropyl, n-butyl,isobutyl and t-butyl.
 35. A compound of claim 32, wherein R₁₅ is takenwith R₁₆ to form a group having the structure:


36. A compound of claim 32, wherein R₁₅ is taken with R₁₆ to form agroup having the structure:


37. A composition comprising an amount of a compound of claim 1effective to modulate GSK3 activity in a human or animal subject whenadministered thereto, together with a pharmaceutically acceptablecarrier.
 38. A composition comprising an amount of a compound of claim20 effective to modulate GSK3 activity in a human or animal subject whenadministered thereto, together with a pharmaceutically acceptablecarrier.
 39. A method of inhibiting GSK3 activity in a human or animalsubject, comprising administering to the human or animal subject in needof such treatment an effective amount of a composition of claim
 37. 40.A method of inhibiting GSK3 activity in a human or animal subject,comprising administering to the human or animal subject in need of suchtreatment an effective amount of a composition of claim
 38. 41. A methodof treating a cell comprising administering to the cell an amount of acompound of claim 1 effective to inhibit GSK3 activity in the cell. 42.A method of treating a cell comprising administering to the cell anamount of a compound of claim 20 effective to inhibit GSK3 activity inthe cell.
 43. A method for treating diabetes or Alzheimer's disease in ahuman or animal subject, comprising administering to the human or animalsubject in need of such treatment an amount of a composition of claim 37effective to inhibit GSK3 activity in the subject.
 44. A method of claim43, wherein the composition is administered by a mode of administrationselected from the group consisting of oral, subcutaneous, transdermal,transmucosal, iontophoretic, intravenous, intrathecal, buccal,sublingual, intranasal, and rectal administration.